Plasmonic assisted systems and methods for interior energy-activation from an exterior source

ABSTRACT

A method and a system for producing a change in a medium disposed in an artificial container. The method places inavicinity of the medium at least one of a plasmonics agent and an energy modulation agent. The method applies an initiation energy through the artificial container to the medium. The initiation energy interacts with the plasmonics agent or the energy modulation agent to directly or indirectly produce the change in the medium. The system includes an initiation energy source configured to apply an initiation energy to the medium to activate the plasmonics agent or the energy modulation agent.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. Ser. No. 15/408,003, filedJan. 17, 2017, now allowed, which is a continuation of U.S. Ser. No.14/851,196 filed Sep. 11, 2015, now U.S. Pat. No/ 9,579,523, which is acontinuation of U.S. Ser. No. 14/245,644 filed Apr. 4, 2014, now U.S.Pat. No. 9,174,190, which is a continuation of U.S. Serial No.13/889,925, filed May 8, 2013, now U.S. Pat. No. 8,927,615, which is acontinuation of U.S. Ser. No. 13/713,931 filed Dec. 13, 2012, now U.S.Pat. No. 9,004,131, which is a continuation of U.S. Ser. No. 12/401,478filed Mar. 10, 2009, now U.S. Pat. No. 8,376,013, the entire contents ofeach of which are hereby incorporated herein by reference. Thisapplication is related to U.S. provisional Ser. No. 60/910,663, filedApr. 8, 2007, entitled “METHOD OF TREATING CELL PROLIFERATIONDISORDERS,” and U.S. non-provisional Ser. No. 11/935,655, filed Nov. 6,2007, entitled “METHOD OF TREATING CELL PROLIFERATION DISORDERS,” thecontents of each of which are hereby incorporated herein by reference.This application is related to U.S. provisional Ser. No. 61/035,559,filed Mar. 11, 2008, entitled “SYSTEMS AND METHODS FOR INTERIORENERGY-ACTIVATION FROM AN EXTERIOR SOURCE,” the entire contents of whichare hereby incorporated herein by reference. This application is relatedto U.S. provisional Ser. No. 61/030,437, filed Feb. 21, 2008, entitled“METHODS AND SYSTEMS FOR TREATING CELL PROLIFERATION DISORDERS USINGPLASMONICS ENHANCED PHOTOSPECTRAL THERAPY (PEPSI) AND EXCITON-PLASMONENHANCED PHOTOTHERAPY (EPEP),” the entire contents of which are herebyincorporated herein by reference. This application is related to U.S.non-provisional Ser. No. 12/389,946, filed Feb. 20, 2009, entitled“METHODS AND SYSTEMS FOR TREATING CELL PROLIFERATION DISORDERS USINGPLASMONICS ENHANCED PHOTOSPECTRAL THERAPY (PEPSI) AND EXCITON-PLASMONENHANCED PHOTOTHERAPY (EPEP),” the entire contents of which are herebyincorporated herein by reference. This application is related to andclaims priority under 35 U.S.C. 119(e) to U.S. provisional Ser. No.61/035,559, filed Mar. 11, 2008, entitled “SYSTEMS AND METHODS FORINTERIOR ENERGY-ACTIVATION FROM AN EXTERIOR SOURCE,” the entire contentsof which are hereby incorporated herein by reference. This applicationis related to and claims priority under 35 U.S.C. 119(e) to U.S.provisional Ser. No. 61/080,140, filed Jul. 11, 2008, entitled“PLASMONIC ASSISTED SYSTEMS AND METHODS FOR INTERIOR ENERGY-ACTIVATIONFROM AN EXTERIOR SOURCE,” the entire contents of which are herebyincorporated herein by reference.

BACKGROUND OF THE INVENTION Field of Invention

The invention relates to methods and systems for generating in theinterior of a medium or body radiant energy for producing a change inthe properties of a medium or body by exposure to the radiation.

Discussion of the Background

Presently, light (i.e., electromagnetic radiation from the radiofrequency through the visible to the x-ray and gamma ray wavelengthrange) activated processing is used in a number of industrial processesranging from photoresist curing, to on-demand ozone production, tosterilization, to the promotion of polymer cross-linking activation(e.g. in adhesive and surface coatings) and others. Today, lightactivated processing is seen in these areas to have distinct advantagesover more conventional approaches. For example, conventionalsterilization by steam autoclaving or in food processing bypasteurization may unsuitably overheat the medium to be sterilized. Assuch, light activated curable coatings are one of the fastest growingsectors in the coatings industry. In recent years, this technology hasmade inroads into a number of market segments like fiber optics, opticaland pressure-sensitive adhesives, and automotive applications like curedtopcoats, and curable powder coatings. The driving force of thisdevelopment is mostly the quest for an increase in productivity of thecoating and curing process, as conventional non light activated adhesiveand surface coatings typically require 1) the elimination of solventsfrom the adhesive and surface coatings to produce a cure and 2) atime/temperature cure which adds delay and costs to the manufacturingprocess.

Moreover, the use of solvent based products in adhesive and surfacecoatings applications is becoming increasingly unattractive because ofrising energy costs and stringent regulation of solvent emissions intothe atmosphere. Optimum energy savings as well as beneficial ecologicalconsiderations are both served by radiation curable adhesive and surfacecoating compositions. Radiation curable polymer cross-linking systemshave been developed to eliminate the need for high oven temperatures andto eliminate the need for expensive solvent recovery systems. In thosesystems, light irradiation initiates free-radical cross-linking in thepresence of common photosensitizers.

However, in the adhesive and surface coating applications and in many ofthe other applications listed above, the light-activated processing islimited due to the penetration depth of light into the processed medium.For example, in water sterilization, ultraviolet light sources arecoupled with agitation and stirring mechanisms in order to ensure thatany bacteria in the water medium will be exposed to the UV light. Inlight-activated adhesive and surface coating processing, the primarylimitation is that the material to be cured must be directly exposed tothe light, both in type (wavelength or spectral distribution) andintensity. In adhesive and surface coating applications, any “shaded”area will require a secondary cure mechanism, increasing cure time overthe non-shaded areas and further delaying cure time due to the existentof a sealed skin through which subsequent curing must proceed (i.e.,referred to as a cocoon effect).

SUMMARY OF THE INVENTION

The invention overcomes the problems and disadvantages of the prior artas described in the various embodiments below.

In one embodiment, there is provided a method and system for producing achange in a medium disposed in an artificial container. The method (1)places in a vicinity of the medium at least one of a plasmonics agentand an energy modulation agent, and (2) applies an initiation energyfrom an applied initiation energy source through the artificialcontainer to the medium. The applied initiation energy interacts withthe plasmonics agent or the energy modulation agent to directly orindirectly produce the change in the medium. The system includes theartificial container configured to contain the medium including theenergy modulation agent or the plasmonics agent. The system furtherincludes an applied initiation energy source configured to apply theinitiation energy through the artificial container to the medium toactivate at least one of the plasmonics agent and the energy modulationagent.

In another embodiment, there is provided a method and system for curinga radiation-curable medium. The method applies an applied energythroughout a composition including an uncured radiation-curable mediumand at least one of a plasmonics agent and an energy modulation agent.The applied initiation energy interacts with the plasmonics agent or theenergy modulation agent to directly or indirectly cure the medium bypolymerization of polymers in the medium. The system includes aninitiation energy source configured to apply initiation energy to thecomposition.

It is to be understood that both the foregoing general description ofthe invention and the following detailed description are exemplary, butare not restrictive of the invention.

BRIEF DESCRIPTION OF THE FIGURES

A more complete appreciation of the invention and many of the attendantadvantages thereof will be readily obtained as the same becomes betterunderstood by reference to the following detailed description whenconsidered in connection with the accompanying drawings, wherein:

FIG. 1 provides an exemplary electromagnetic spectrum in meters (1 nmequals 10⁻⁹ meters);

FIG. 2 is a table providing a list of photoactivatable agents;

FIG. 3A is a schematic depicting a system according to one embodiment ofthe invention in which an initiation energy source is directed to aself-contained medium for producing changes in the medium;

FIG. 3B is a schematic depicting a system according to anotherembodiment of the invention in which the initiation energy source isdirected to a container enclosing a medium having energy modulationagents disbursed within the medium;

FIG. 3C is a schematic depicting a system according to anotherembodiment of the invention in which the initiation energy source isdirected to a container enclosing a medium having energy modulationagents segregated within the medium;

FIG. 3D is a schematic depicting a system according to anotherembodiment of the invention in which the initiation energy source isdirected to a container enclosing a medium having energy modulationagents segregated within the medium in a fluidized bed configuration;

FIG. 4 illustrates an exemplary computer system for implementing variousembodiments of the invention;

FIGS. 5A and 5B are representations of plasmonic nanostructures andtheir theoretical electromagnetic enhancement at different excitationwavelength;

FIGS. 6A-6G provide representative embodiments of plasmonicsphoto-active probes useful in the invention;

FIGS. 7A and 7B are graphical explanations of the plasmonics-enhancedeffect of the invention;

FIGS. 8A-8J show representative embodiments of plasmonics-activenanostructures;

FIGS. 9A-9C are representations of several embodiments of PEPST probeswith a linker that can be cut by a photon radiation;

FIG. 10 is a representation of the “window” in hydrous medium;

FIG. 11 is a representation of an embodiment of the energy modulationagent (or excitation energy converter/EEC)-photo activator (PA) systemof the invention;

FIGS. 12A-12F are representations of several embodiments of plasmonicsphoto-active energy modulation agent-PA probes;

FIGS. 13A-13B show structures of various preferred embodiments of goldcomplexes exhibiting XEOL;

FIG. 14 shows the structure of a further embodiment of compoundexhibiting XEOL, namely a tris-8-hydroxyquinoline-aluminum complex;

FIG. 15 is a representation of a plasmonics-enhanced mechanism for aphoto-active energy modulation agent-PA probe of the invention;

FIGS. 16A-16C are representations of embodimens of a PEPST energymodulation agent-PA system with detachable bond;

FIG. 17 is a representation of an embodiment of PEPST probes for dualplasmonic excitation;

FIGS. 18A-18D provides a representation of the sequence for use ofencapsulated photoactive agents;

FIG. 19 is a graph showing the XEOL of Eu doped in BaFBr matrix;

FIG. 20 shows various embodiments of EIP probes of the invention;

FIG. 21A-21B show further embodiments of EIP probes of the invention;

FIG. 22A-22C show further embodiments of schematic designs of HP probes;

FIG. 23A and 23B are representations of various embodiments of basicEPEP probes;

FIG. 24 is a representation of one embodiment of EPEP probes having NPs,NWs and

FIG. 25 is a representation of one embodiment of EPEP probes having NPs,NWs NRs and bioreceptors;

FIG. 26 is a representation of an embodiment of EPEP probes having NPsand multiple NWs;

FIG. 27 is a representation of an embodiment of a sterilization systemof the invention;

FIG. 28 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes plasmonics;

FIG. 29 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes a photo-active material;

FIG. 30 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes a photo-active material and adielectric medium;

FIG. 31 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes an X-ray energy converter withembedded metal nanoparticles serving as a plasmonics function;

FIG. 32 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes embedded metal nanoparticlesincorporated on re-entrant structures inside where a medium to besterilized will flow;

FIG. 33 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes an X-ray energy converter with theembedded metal nanoparticles of FIG. 31 included on an inside layer of acontainer where a medium to be sterilized will flow;

FIG. 34 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes embedded metal nanoparticlesincorporated on re-entrant wall structures inside where a medium to besterilized will flow;

FIG. 35 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes chemical receptors on an inside ofa container where a medium to be sterilized will flow;

FIG. 36 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes embedded metal nanoparticles inone layer and chemical receptors in another more interior layer on aninside of a container where a medium to be sterilized will flow;

FIG. 37 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes a photo-active material andchemical receptors on an inside of a container where a medium to besterilized will flow;

FIG. 38 is a representation of another embodiment of a sterilizationsystem of the invention that utilizes a photo-active material, adielectric layer in conjunction with embedded metal nanoparticles, andchemical receptors on a surface of the probe inside of a container wherea medium to be sterilized will flow;

FIG. 39 is a representation of an embodiment of a sterilization probesystem of the invention;

FIG. 40 is a representation of another embodiment of a sterilizationprobe system of the invention that utilizes a dielectric layer nconjunction with embedded metal nanoparticles,

FIG. 41 is a representation of another embodiment of a sterilizationprobe system of the invention that utilizes an X-ray energy converterand chemical receptors on a surface of the probe inside of a containerwhere a medium to be sterilized will flow;

FIG. 42 is a representation of another embodiment of a sterilizationprobe system of the invention that utilizes an X-ray energy converterand further a dielectric layer in conjunction with embedded metalnanoparticles on a surface of the probe inside of a container where amedium to be sterilized will flow;

FIGS. 43A-43D are representations of another embodiment of asterilization system of the invention that utilizes paramagnetic corematerials; and

FIG. 44A-44G are representations of different plasmonics probes of theinvention.

DETAILED DESCRIPTION OF THE INVENTION

The invention sets forth a novel method for causing a change in activityof an in a medium that is effective, specific, and able to produce achange to the medium.

Generally, the invention provides methods for producing a change in amedium after generation of radiant light inside the medium. In thismethod, an initiation energy source provides an initiation energy thatpenetrates the medium and induces internal radiation to produce adesired effect in the medium.

In one embodiment, the initiation energy source is applied directly orindirectly to the medium. Within the context of the invention, thephrase “applied indirectly” (or variants of this phrase, such as“applying indirectly”, “indirectly applies”, “indirectly applied”,“indirectly applying”, etc.), when referring to the application of theinitiation energy, means the penetration by the initiation energy intothe medium beneath the surface of the medium and to the activatableagenst or energy modulation agents within a medium. In one embodiment,the initiation energy interacts with a previously supplied energymodulation agent which then activates the activatable agent.

Although not intending to be bound by any particular theory or beotherwise limited in any way, the following theoretical discussion ofscientific principles and definitions are provided to help the readergain an understanding and appreciation of the invention.

As used herein, an “activatable agent” is an agent that normally existsin an inactive state in the absence of an activation signal. When theagent is activated by an activation signal under activating conditions,the agent is capable of producing a desired pharmacological, cellular,chemical, electrical, or mechanical effect in a medium (i.e. apredetermined change). For example, when photocatalytic agents areirradiated with visible or UV light, these agents induce polymerizationand “curing” of light sensitive adhesives.

Signals that may be used to activate a corresponding agent may include,but are not limited to, photons of specific wavelengths (e.g. x-rays, orvisible light), electromagnetic energy (e.g. radio or microwave),thermal energy, acoustic energy, or any combination thereof. Activationof the agent may be as simple as delivering the signal to the agent ormay further require a set of activation conditions. For example, anactivatable agent, such as a photosensitizer, may be activated by UV-Aradiation (e.g., by UV-A radiation generated internally in the medium).Once activated, the agent in its active-state may then directly proceedto produce a predetermined change.

Where activation may further require other conditions, mere delivery ofthe activation signal may not be sufficient to bring about thepredetermined change. For example, a photoactive compound that achievesits effect by binding to certain structure in its active state mayrequire physical proximity to the target structure when the activationsignal is delivered. For such activatable agents, delivery of theactivation signal under non-activating conditions will not result in thedesired effect. Some examples of activating conditions may include, butare not limited to, temperature, pH, location, state of the medium, andthe presence or absence of co-factors.

Selection of an activatable agent greatly depends on a number of factorssuch as the desired change, the desired form of activation, as well asthe physical and biochemical constraints that may apply. Exemplaryactivatable agents may include, but are not limited to agents that maybe activated by photonic energy, electromagnetic energy, acousticenergy, chemical or enzymatic reactions, thermal energy, microwaveenergy, or any other suitable activation mechanisms.

When activated, the activatable agent may effect changes that include,but are not limited to an increase in organism activity, afelnientation, a decrease in organism activity, apoptosis, redirectionof metabolic pathways, a sterilization of a medium, a crosspolymerization and curing of a medium, or a cold pasteurization of amedium.

The mechanisms by which an activatable agent may achieve its desiredeffect are not particularly limited. Such mechanisms may include directaction on a predetermined target as well as indirect actions viaalterations to the biochemical pathways. In one embodiment, theactivatable agent is capable of chemically binding to the organism in amedium. In this embodiment, the activatable agent, is exposed in situ toan activating energy emitted from an energy modulation agent, which, inturn receives energy from an initiation energy source.

Suitable activatable agents include, but are not limited to, photoactiveagents, sono-active agents, thermo-active agents, andradio/microwave-active agents. An activatable agent may be a smallmolecule; a biological molecule such as a protein, a nucleic acid orlipid; a supramolecular assembly; a nanoparticle; or any other molecularentity capable of producing a predetermined activity once activated.

The activatable agent may be derived from a natural or synthetic origin.Any such molecular entity that may be activated by a suitable activationsignal source to effect a predetermined cellular change may beadvantageously employed in the invention.

Suitable photoactive agents include, but are not limited to: psoralensand psoralen derivatives, pyrene cholesteryloleate, acridine, porphyrin,fluorescein, rhodamine, 16-diazorcortisone, ethidium, transitionansition metal complexes of bleomycin, transition metal complexes ofdeglycobleomycin, organoplatinum complexes, alloxazines such as7,8-dimethyl-10-ribityl isoalloxazine (riboflavin), 7,8,10-triethylisoalloxazine (lumiflavin), 7,8-dimethylalloxazine (lumichrome),isoalloxazine-adenine dinucleotide (flavine adenine dinucleotide [FAD]),alloxazine mononucleotide (also known as flavine mononucleotide [FMN]and riboflavine-5-phosphate), vitamin Ks, vitamin L, their metabolitesand precursors, and napththoquinones, naphthalenes, naphthols and theirderivatives having planar molecular conformations, porphyrins, dyes suchas neutral red, methylene blue, acridine, toluidines, flavine(acriflavine hydrochloride) and phenothiazine derivatives, coumarins,quinolones, quinones, and anthroquinones, aluminum (111) phthalocyaninetetrasulfonate, hematoporphyrin, and phthalocyanine, and compounds whichpreferentially adsorb to nucleic acids with little or no effect onproteins. The term “alloxazine” includes isoalloxazines.

Endogenously-based derivatives include synthetically derived analogs andhomologs of endogenous photoactivated molecules, which may have or lacklower (1 to 5 carbons) alkyl or halogen substitutes of thephotosensitizers from which they are derived, and which preserve thefunction and substantial non-toxicity. Endogenous molecules areinherently non-toxic and may not yield toxic photoproducts afterphotoradiation.

FIG. 1 provides an exemplary electromagnetic spectrum in meters (1 nmequals 1 nanometer). As used herein, an “energy modulation agent” refersto an agent that is capable of receiving an energy input from a sourceand then re-emitting a different energy to a receiving target. Energytransfer among molecules may occur n a number of ways. The form ofenergy may be electronic, thermal, electromagnetic, kinetic, or chemicalin nature. Energy may be transferred from one molecule to another(intermolecular transfer) or from one part of a molecule to another partof the same molecule (intramolecular transfer). For example, amodulation agent may receive electromagnetic energy and re-emit theenergy in the form of thermal energy.

Table 1 in FIG. 2 provides a list of photoactivatable agents that may beused as primary or secondary internal light sources. For example, thephotoactivatable agents could be receptors of X-ray induced emissionsfrom nanoparticles (to be discussed later) and which in turn emit asecondary light. In some mediums, it may be that the excitationwavelengths in Table 1 are transparent to the particular medium and theemission wavelengths are highly absorbent (due to, for example,molecular or solid state band gap transitions). In those cases, thephotoreactive agents in Table 1 would be the primary sources forinternal light generation.

In various embodiments, the energy modulation agent receives higherenergy (e.g. x-ray) and re-emits in lower energy (e.g. UV-A). Somemodulation agents may have a very short energy retention time (on theorder of fs, e.g. fluorescent molecules) whereas others may have a verylong half-life (on the order of minutes to hours, e.g. luminescent orphosphorescent molecules). Suitable energy modulation agents include,but are not limited to, a biocompatible fluorescing metal nanoparticle,fluorescing dye molecule, gold nanoparticle, a water soluble quantum dotencapsulated by polyamidoamine dendrimers, a luciferase, a biocompatiblephosphorescent molecule, a combined electromagnetic energy harvestermolecule, and a lanthanide chelate capable of intense luminescence.Typically, the energy modulation agents induce photoreactive changes inthe medium and are not used for the purpose of exclusively heating themedium.

Various exemplary uses are described in the embodiments below.

The modulation agents may further be coupled to a carrier for targetingpurposes. For example, a biocompatible molecule, such as a fluorescingmetal nanoparticle or fluorescing dye molecule that emits in the UV-Aband, may be selected as the energy modulation agent. The energymodulation agent may be preferably directed to the desired site bysystemic administration into a medium. For example, a UV-A emittingenergy modulation agent may be distributed in the medium by physicalinsertion and or mixing, or by conjugating the UV-A emitting energymodulation agent with a specific carrier, such as a lipid, chitin orchitin-derivative, a chelate or other functionalized carrier that iscapable of concentrating the UV-A emitting source in a specific targetregion of the medium.

Additionally, the energy modulation agent can be used alone or as aseries of two or more energy modulation agents such that the energymodulation agents provide an energy cascade. Thus, the first energymodulation agent in the cascade will absorb the activation energy,convert it to a different energy which is then absorbed by the secondenergy modulation in the cascade, and so forth until the end of thecascade is reached with the final energy modulation agent in the cascadeemitting the energy necessary to activate the activatable agent.Alternatively, one or more energy modulation agents in the cascade mayalso activate additional activatable agents.

Although the activatable agent and the energy modulation agent can bedistinct and separate, it will be understood that the two agents neednot be independent and separate entities. In fact, the two agents may beassociated with each other via a number of different configurations.Where the two agents are independent and separately movable from eachother, they can generally interact with each other via diffusion andchance encounters within a common surrounding medium. Where theactivatable agent and the energy modulation agent are not separate, theymay be combined into one single entity.

The initiation energy source can be any energy source capable ofproviding energy at a level sufficient to activate the activatable agentdirectly, or to provide the energy modulation agent with the inputneeded to emit the activation energy for the activatable agent (indirectactivation). Preferable initiation energy sources include, but are notlimited to, ultraviolet lamps such as UV-A and UV-B lamps, halogenlamps, fiber optic lines, a light needle, an endoscope, self-ballastedmercury vapor lamps, ballasted HID lamps, and any device capable ofgenerating x-ray, y-ray, gamma-ray, or electron beams.

In one embodiment, the initiation energy is capable of penetratingcompletely through the medium. Within the context of the invention, thephrase “capable of penetrating completely through the medium” is used torefer to energy capable of penetrating a container to any distancenecessary to activate the activatable agent within the medium. It is notrequired that the energy applied actually pass completely through themedium, merely that it be capable of doing so in order to permitpenetration to any desired distance to activate the activatable agent.The type of energy source chosen will depend on the medium itself.Exemplary initiation energy sources that are capable of penetratingcompletely through the medium include, but are not limited to, x-rays,gamma rays, electron beams, microwaves and radio waves.

In one embodiment, the source of the initiation energy can be aradiowave emitting nanotube, such as those described by K. Jensen, J.Weldon, H. Garcia, and A. Zettl in the Department of Physics at theUniversity of California at Berkeley (seehttp://socrates.berkeley.edu/˜argon/nanoradio/radio.html, the entirecontents of which are hereby incorporated by reference). These nanotubescan be introduced to the medium, and preferably would be coupled to theactivatable agent or the energy modulation agent, or both, such thatupon application of the initiation energy, the nanotubes would acceptthe initiation energy (preferably radiowaves), then emit radiowaves inclose proximity to the activatable agent, or in close proximity to theenergy modulation agent, to then cause activation of the activatableagent. In such an embodiment, the nanotubes would act essentially as aradiowave focusing or amplification device in close proximity to theactivatable agent or energy modulation agent.

Alternatively, the energy emitting source may be an energy modulationagent that emits energy in a form in suitable for absorption by atransfer agent or for direct interaction with components of the medium.For example, the initiation energy source may be acoustic energy, andone energy modulation agent may be capable of receiving acoustic energyand emitting photonic energy (e.g. sonoluminescent molecules) to bereceived by another energy modulation agent that is capable of receivingphotonic energy. Other examples include transfer agents that receiveenergy at x-ray wavelength and emit energy at UV wavelength, preferablyat UV-A wavelength. As noted above, a plurality of such energymodulation agents may be used to form a cascade to transfer energy frominitiation energy source via a series of energy modulation agents toactivate the activatable agent.

Photoactivatable agents may be stimulated by an energy source throughmechanisms such as irradiation, resonance energy transfer, excitonmigration, electron injection, or chemical reaction, to an activatedenergy state that is capable of producing the predetermined changedesired. One advantage is that wavelengths of emitted radiation may beused to selectively stimulate one or more photoactivatable agents orenergy modulation agents capable of stimulating the one or morephotoactivatable agents. The energy modulation agent is suitablystimulated at a wavelength and energy that causes little or no change tothe medium.

In another embodiment, he photoactivatable agent is stimulated via aresonance energy transfer. Resonance Energy Transfer (RET) is an energytransfer mechanism between two molecules having overlapping emission andabsorption bands. Electromagnetic emitters are capable of converting anarriving wavelength to a longer wavelength. For example, UV-B energyabsorbed by a first molecule may be transferred by a dipole-dipoleinteraction to a UV-A-emitting molecule in close proximity to theUV-B-absorbing molecule. One advantage is that multiple wavelengths ofemitted radiation may be used to selectively stimulate one or morephotoactivatable agents or energy modulation agents capable ofstimulating the one or more photoactivatable agents. With RET, theenergy modulation agent is preferably stimulated at a wavelength andenergy that causes little or no effect to the surrounding medium withthe energy from one or more energy modulation agents being transferred,such as by Foerster Resonance Energy Transfer, to the photoactivatableagents.

Alternatively, a material absorbing a shorter wavelength may be chosento provide RET to a non-emitting molecule that has an overlappingabsorption band with the transferring molecule's emission band.Alternatively, phosphorescence, chemiluminescence, or bioluminescencemay be used to transfer energy to a photoactivatable molecule.

Alternatively, one can apply the initiation energy source to the medium.Within the context of the invention, the applying of the initiationenergy source means the application of an agent, that itself producesthe initiation energy, in a manner that permits the agent to arrive atthe target structure within the medium. The application can take anyform. Further, the initiation energy source in this embodiment can be inany form, including, but not limited to, tablet, powder, liquidsolution, liquid suspension, liquid dispersion, gas or vapor, etc. Inthis embodiment, the initiation energy source includes, but is notlimited to, chemical energy sources, nanoemitters, nanochips, and othernanomachines that produce and emit energy of a desired frequency.

Recent advances in nanotechnology have provided examples of variousdevices that are nanoscale and produce or emit energy, such as theMolecular Switch (or Mol-Switch) work by Dr. Keith Firman of the ECResearch and Development Project, or the work of Cornell et al. (1997)who describe the construction of nanomachines based around ion-channelswitches only 1.5 nm in size, which use ion channels formed in anartificial membrane by two gramicidin molecules: one in the lower layerof the membrane attached to a gold electrode and one in the upper layertethered to biological receptors such as antibodies or nucleotides. Whenthe receptor captures a target molecule or cell, the ion channel isbroken, its conductivity drops, and the biochemical signal is convertedinto an electrical signal. These nanodevices could also be coupled withthe invention to provide targeting of the target cell, to deliver theinitiation energy source directly at the desired site.

In another embodiment, the invention includes the application of theactivatable agent, along with a source of chemical energy such aschemiluminescence, phosphorescence or bioluminescence. The source ofchemical energy can be a chemical reaction between two or morecompounds, or can be induced by activating a chemiluminescent,phosphorescent or bioluminescent compound with an appropriate activationenergy, either outside the medium or inside the medium, with thechemiluminescence, phosphorescence or bioluminescence being allowed toactivate the activatable agent in the medium. The administration of theactivatable agent and the source of chemical energy can be performedsequentially in any order or can be performed simultaneously.

In the case of certain sources of such chemical energy, the applicationof the chemical energy source can be performed after activation outsidethe medium, with the lifetime of the emission of the energy being up toseveral hours for certain types of phosphorescent materials for example.

When molecules absorb excitation light, electrons undergo transitionsfrom the ground state to an excited electronic state. The electronicexcitation energy subsequently relaxes via radiative emission(luminescence) and radiationless decay channels. When a molecule absorbsexcitation energy, it is elevated from S_(o) to some vibrational levelof one of the excited singlet states, S_(n), in the manifold S₁, . . . ,S_(n). In condensed media (tissue), the molecules in the S_(n) statedeactivate rapidly, within 10⁻¹³ to 10^(—11) s via vibrationalrelaxation (VR) processes, ensuring that they are in the lowestvibrational levels of S_(n) possible. Since the VR process is fasterthan electronic transitions, any excess vibrational energy is rapidlylost as the molecules are deactivated to lower vibronic levels of thecorresponding excited electronic state. This excess VR energy isreleased as thermal energy to the surrounding medium. From the S_(n)state, the molecule deactivates rapidly to the isoenergetic vibrationallevel of a lower electronic state such as S_(n−1) via an internalconversion (IC) process. IC processes are transitions between states ofthe same multiplicity.

The molecule subsequently deactivates to the lowest vibronic levels ofS_(n−1) via a VR process. By a succession of IC processes immediatelyfollowed by VR processes, the molecule deactivates rapidly to the groundstate S₁. This process results in excess VR and IC energy released asthermal energy to the surrounding medium leading to the overheating ofthe local environment surrounding the light absorbing drug molecules.The heat produced results in local changes in the medium.

Light absorbing species in various embodiments can include naturalchromophores in tissue or exogenous dye compounds such as indocyaninegreen, naphthalocyanines, and porphyrins coordinated with transitionmetals and metallic nanoparticles and nanoshells of metals. Naturalchromophores, however, suffer from very low absorption. The choice ofthe exogenous photothermal agents is made on the basis of their strongabsorption cross sections and highly efficient light-to-heat conversion.This feature greatly minimizes the amount of laser energy needed toinduce a local change in the medium.

One problem associated with the use of dye molecules is theirphotobleaching under laser irradiation. Therefore, nanoparticles such asgold nanoparticles and nanoshells have recently been used. The promisingrole of nanoshells in medical applications has been demonstrated[Hirsch, L. R., Stafford , R. J., Bankson, J. A. , Sershen, S. R.,Rivera, B., Price, R. E., Hazle, J. D., Halas, N. J., and West, J. L.,Nanoshell-mediated near-infrared thermal therapy of tumors undermagnetic resonance guidance. PNAS, 2003. 100(23): p. 13549-13554], theentire contents of which are incorporated herein by reference. The useof plasmonics-enhanced photothermal properties of metal nanoparticlesfor photothermal therapy has also been reviewed (Xiaohua Huang &Prashant K. Jain & Ivan H. El-Sayed & Mostafa A. El-Sayed, “Plasmonicphotothermal therapy (PPTT) using gold nanoparticles ” Lasers in MedicalScience, August 2007), the entire contents of which are incorporatedherein by reference.

Yet another example is that nanoparticles or nanoclusters of certainatoms may be introduced such that they are capable of resonance energytransfer over comparatively large distances, such as greater than onenanometer, more preferably greater than five nanometers, even morepreferably at least 10 nanometers. Functionally, resonance energytransfer may have a large enough “Foerster” distance (R₀), such thatnanoparticles in one part of a medium are capable of stimulatingactivation of photoactivatable agents disposed in a distant portion ofthe medium, so long as the distance does not greatly exceed R₀. Forexample, gold nanospheres having a size of 5 atoms of gold have beenshown to have an emission band in the ultraviolet range, recently.

Any of the photoactivatable agents may be exposed to an excitationenergy source provided in the medium. The photoactive agent may bedirected to a receptor site by a carrier having a strong affinity forthe receptor site. Within the context of the invention, a “strongaffinity” is preferably an affinity having an equilibrium dissociationconstant, K_(i), at least in the nanomolar, nM, range or higher. Thecarrier may be a polypeptide and may form a covalent bond with aphotoactive agent, for example. Alternatively, a photoactive agent mayhave a strong affinity for the target molecule in the medium withoutbinding to a carrier.

In one embodiment, a plurality of sources for supplying electromagneticradiation energy or energy transfer is provided by one or more moleculesprovided to the medium. The molecules may emit stimulating radiation inthe correct band of wavelength to stimulate the photoactivatable agents,or the molecules may transfer energy by a resonance energy transfer orother mechanism directly to the photoactivatable agent or indirectly bya cascade effect via other molecular interactions.

In a further embodiment, a biocompatible emitting source, such as afluorescing metal nanoparticle or fluorescing dye molecule, is selectedthat emits in the UV-A band. UV-A and the other UV bands are known to beeffective as get inicides.

In one embodiment, the UV-A emitting source is a gold nanoparticlecomprising a cluster of 5 gold atoms, such as a water soluble quantumdot encapsulated by polyamidoamine dendrimers. The gold atom clustersmay be produced through a slow reduction of gold salts (e.g. HAuCl₄ orAuBr₃) or other encapsulating amines, for example. One advantage of sucha gold nanoparticle is the increased Foerster distance (i.e. R₀), whichmay be greater than 100 angstroms. The equation for determining theFoerster distance is substantially different from that for molecularfluorescence, which is limited to use at distances less than 100angstroms. It is believed that the gold nanoparticles are governed bynanoparticle surface to dipole equations with a 1/R⁴ distance dependencerather than a 1/R⁶ distance dependence. For example, this permitscytoplasmic to nuclear energy transfer between metal nanoparticles and aphotoactivatable molecule.

In another embodiment, a UV or light-emitting luciferase is selected asthe emitting source for exciting a photoactivatable agent. A luciferasemay be combined with molecules, which may then be oxygenated withadditional molecules to stimulate light emission at a desiredwavelength. Alternatively, a phosphorescent emitting source may be used.Phosphorescent materials may have longer relaxation times thanfluorescent materials, because relaxation of a triplet state is subjectto forbidden energy state transitions, storing the energy in the excitedtriplet state with only a limited number of quantum mechanical energytransfer processes available for returning to the lower energy state.Energy emission is delayed or prolonged from a fraction of a second toseveral hours. Otherwise, the energy emitted during phosphorescentrelaxation is not otherwise different than fluorescence, and the rangeof wavelengths may be selected by choosing a particular phosphor.

In another embodiment, a combined electromagnetic energy harvestermolecule is designed, such as the combined light harvester disclosed inJ. Am. Chem. Soc. 2005, 127, 9760-9768, the entire contents of which arehereby incorporated by reference. By combining a group of fluorescentmolecules in a molecular structure, a resonance energy transfer cascademay be used to harvest a wide band of electromagnetic radiationresulting in emission of a narrow band of fluorescent energy. By pairinga combined energy harvester with a photoactivatable molecule, a furtherenergy resonance transfer excites the photoactivatable molecule, whenthe photoactivatable molecule is nearby stimulated combined energyharvester molecules. Another example of a harvester molecule isdisclosed in FIG. 4 of “Singlet-Singlet and Triplet-Triplet EnergyTransfer in Bichromophoric Cyclic Peptides,” M. S. Thesis by M. O.Guler, Worcester Polytechnic Institute, May 18, 2002, which isincorporated herein by reference.

In another embodiment, a Stokes shift of an emitting source or a seriesof emitting sources arranged in a cascade is selected to convert ashorter wavelength energy, such as X-rays, to a longer wavelengthfluorescence emission such a optical or UV-A, which is used to stimulatea photoactivatable molecule in the medium.

In an additional embodiment, the photoactivatable agent can be aphotocaged complex having an active agent (which can be a cytotoxicagent if cytotoxicity is needed, or can be an activatable agent)contained within a photocage. In various embodiments, where the activeagent is a cyotoxic agent, the photocage molecule releases the cytotoxicagent into the medium where it can attack non-beneficial “target”species in the medium. The active agent can be bulked up with othermolecules that prevent it from binding to specific targets, thus maskingits activity. When the photocage complex is photoactivated, the bulkfalls off, exposing the active agent. In such a photocage complex, thephotocage molecules can be photoactive (i.e. when photoactivated, theyare caused to dissociate from the photocage complex, thus exposing theactive agent within), or the active agent can be the photoactivatableagent (which when photoactivated causes the photocage to fall off), orboth the photocage and the active agent are photoactivated, with thesame or different wavelengths. Suitable photocages include thosedisclosed by Young and Deiters in “Photochemical Control of BiologicalProcesses”, Org. Biomol. Chen , 5, pp. 999 - 1005 (2007) and“Photochemical Hammerhead Ribozyme Activation”, Bioorganic & MedicinalChemistry Letters, 16(10) ,pp. 2658-2661 (2006), the contents of whichare hereby incorporated by reference.

Work has shown that the amount of singlet oxygen necessary to cause celllysis, and thus cell death, is 0.32 H 10⁻³ mol/liter or more, or 10⁹singlet oxygen molecules/cell or more. In one embodiment of theinvention, the level of singlet oxygen production caused by theinitiation energy or the activatable agent upon activation is sufficientto cause a change in a medium, wherein the medium becomes free from anymicroorganisms. Microorganisms include but are not limited to bacteria,viruses, yeasts or fungi. To this end, singlet oxygen in sufficientamounts as described above can be used to sterilize the medium.

For example, medical bottle caps need to be sterilized between the basecap material and the glued seal material which contacts the base of themedical bottle. Because steam autoclaves are insufficient for thispurpose, one embodiment of the invention uses UV luminescing particlesincluded in the adhesive layer when the seal material is applied to thebottle cap. Then, X-ray irradiation becomes capable of curing theadhesive and producing within the adhesive medium UV radiation fordirect sterilization or the production of singlet oxygen or ozone forbiological germicide.

The activatable agent and derivatives thereof as well as the energymodulation agent, can be incorporated into compositions suitable fordelivery to particular mediums. The composition can also include atleast one additive having a complementary effect upon the medium, suchas a lubricant or a sealant.

The carrier can be a solvent or dispersion medium containing, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyethylene glycol, and the like), and suitablemixtures thereof. The proper fluidity can be maintained, for example, bythe use of a coating such as lecithin, by the maintenance of therequired particle size in the case of dispersion and by the use ofsurfactants.

Referring to FIG. 3A, an exemplary system according to one embodiment ofthe invention may have an initiation energy source 1 directed at medium4. Activatable agents 2 and an energy modulation agents 3 are dispersedthroughout the medium 4. The initiation energy source 1 may additionallybe connected via a network 8 to a computer system 5 capable of directingthe delivery of the initiation energy. In various embodiments, theenergy modulation agents 3 are encapsulated energy modulation agents 6,depicted in FIG. 3A as silica encased energy modulation agents. As shownin FIG. 3A, initiation energy 7 in the form of radiation from theinitiation energy source 1 permeated throughout the medium 4. A morethorough discussion of the computer system 5 is provided below inreference to FIG. 4. As discussed below in more detail, the initiationenergy source 1 can be an external energy source or an energy sourcelocated at least partially in the medium 4. As discussed below in moredetail, activatable agents 2 and/or the energy modulation agents 3 caninclude plasmonics agents which enhance either the applied energy or theenergy emitted from the energy modulation agents 3 so as to directly orindirectly produce a change in the medium.

In various embodiments, the initiation energy source 1 may be a linearaccelerator equipped with image guided computer-control capability todeliver a precisely calibrated beam of radiation to a pre-selectedcoordinate. One example of such linear accelerators is the SmartBeam™IMRT (intensity modulated radiation therapy) system from Varian medicalsystems (Varian Medical Systems, Inc., Palo Alto, Calif.). In otherembodiments, the initiation energy source 1 may be commerciallyavailable components of X-ray machines or non-medical X-ray machines.X-ray machines that produce from 10 to 150 keV X-rays are readilyavailable in the marketplace. For instance, the General ElectricDefinium series or the Siemens MULTIX series are but two examples oftypical X-ray machines designed for the medical industry, while theEagle Pack series from Smith Detection is an example of a non-medicalX-ray machine. As such, the invention is capable of performing itsdesired function when used in conjunction with commercial X-rayequipment.

In other embodiments, the initiation energy source 1 can be a radiofrequency or microwave source emitting radio waves at a frequency whichpermeates the medium and which triggers or produces secondary radiantenergy emission within the medium by interaction with the energymodulation elements 6 therein. In other embodiments, the initiationenergy source 1 can be an ultraviolet, visible, near infrared (NIR) orinfrared (IR) emitter emitting at a frequency which permeates the medium4 and which triggers or produces secondary radiant energy emissionwithin medium 4 by interaction with the energy modulation elements 6therein.

FIG. 3B is a schematic depicting another system according to anotherembodiment of the invention in which the initiation energy source 1 ofFIG. 3A is directed to energy modulation elements 6 placed in thevicinity of a fluid medium 4 (e.g., a liquid or other fluid-like medium)and held inside a container 9. The container 9 is made of a materialthat is “transparent” to the radiation 7. For example, plastic, quartz,glass, or aluminum containers would be sufficiently transparent toX-rays, while plastic or quartz or glass containers would be transparentto microwave or radio frequency light. The energy modulation elements 6can be dispersed uniformly throughout the medium or may be segregated indistinct parts of the medium or further separated physically from themedium by encapsulation structures 10. A supply 11 provides the medium 4to the container 9.

Alternatively, as shown in FIG. 3C, the luminescing particles could bepresent in the medium in encapsulated structures 10. In one embodiment,the encapsulated structures 10 are aligned with an orientation in linewith the external initiation energy source 1. In this configuration,each of the encapsulated structures 10 has itself a “line-of-sight” tothe external initiation energy source 1 shown in FIG. 3C without beingoccluded by other of the encapsulated structures 10. In otherembodiments, the encapsulated structures 10 are not so aligned in thatdirection, but could aligned perpendicular to the direction shown inFIG. 3C, or could be randomly placed. Indeed, supply of fluid medium 4could itself be used to agitate the encapsulated structures 10 and mixthe fluid medium 4 inside container 9.

The system of FIG. 3C may also be used without energy modulation agents.In this embodiment, the initiation energy source 1 can be for example atan energy suitable for driving physical, chemical, and/or biologicalprocesses in the fluid medium 4. The plasmonics agents included in theencapsulated structures 10 effectively amplify the light from theinitiation energy source 1 as it interacts with the medium 4. In oneaspect of the invention, the initiation energy source 1 can a UV lightsource as in many conventional UV sterilization systems and theencapsulated structures 10 of FIG. 3C are light rods conducting UV lightfrom an exterior source to a region inside the medium 4. In one aspectof the invention, the initiation energy source 1 can be even disposedinside the medium and can be a UV light source as in many conventionalUV sterilization systems.

FIG. 3D is a schematic depicting a system according to anotherembodiment of the invention in which the initiation energy source isdirected a container enclosing a medium having energy modulation agentssegregated within the medium in a fluidized bed 20 configuration. Thefluidized bed 20 includes the encapsulated structures 10 in aconfiguration where a fluid to be treated is passed between theencapsulated structures 10. The encapsulated structures 10 can includeboth energy modulation agents and plasmonics agents as described herein.

In further embodiments of the invention, robotic manipulation devicesmay also be included in the systems of FIG. 3A, 3B, 3C, and 3D for thepurpose of delivering and dispersing the energy modulation elements 6 inmedium 4 or for the purpose of removing old product and introducing newproduct for treatment into the system.

In the invention, energy transfer among molecules may occur in a numberof ways. The foi in of energy may be electronic, thermal,electromagnetic, kinetic, or chemical in nature. The energy can bemodulated up to emit higher energy from the energy modulation agentcompared to the input initiation energy, or can be modulated down toemit lower energy from the energy modulation agent compared to the inputinitiation energy. Energy may be transferred from one molecule toanother (intermolecular transfer) or from one part of a molecule toanother part of the same molecule (intramolecular transfer). Forexample, a modulation agent may receive electromagnetic energy andre-emit the energy in the form of a different energy. In variousembodiments, the energy modulation agents receive higher energy (e.g.x-ray) and re-emits in lower energy (e.g. UV-A). In other embodiments,the energy modulation agent receive lower energy (e.g., infrared ornear-infrared) and emits in a higher energy (e.g., visible orultraviolet). Energy transfer processes are also referred to asmolecular excitation. Some modulation agents may have a very shortenergy retention time (on the order of fs-ns, e.g. fluorescentmolecules) whereas others may have a very long half-life (on the orderof seconds to hours, e.g. luminescent inorganic molecules orphosphorescent molecules). Suitable energy modulation agents include,but are not limited to, a metal nanoparticle or a biocompatible metalnanoparticle, a metal coated or uncoated with a biocompatible outerlayer, a chemiluminescent molecule whose rate of luminescence isincreased by microwave activation, fluorescing dye molecule, goldnanoparticle, a water soluble quantum dot encapsulated by polyamidoaminedendrimers, a luciferase, a biocompatible phosphorescent molecule, abiocompatible fluorescent molecule, a biocompatible scattering molecule,a combined electromagnetic energy harvester molecule, and a lanthanidechelate capable of intense luminescence. Various exemplary uses of theseare described.

The modulation agents may further be coupled to a carrier for targetingpurposes. For example, a biocompatible molecule, such as a fluorescingmetal nanoparticle or fluorescing dye molecule that emits in the UV-Aband, may be selected as the energy modulation agent.

The energy modulation agent may be preferably directed to the desiredsite (e.g. in close vicinity to a photoactive substance such as forexample a photocatalyst or a photo initiator) by pre-distribution of theenergy modulation agent into a medium to be exposed to the activationenergy. For example, a UV-A emitting energy modulation agent may beconcentrated in joints for adhesion of two parts together by physicalinsertion or by conjugating the UV-A emitting energy modulation agentwith a photoactivatable resin.

Additionally, the energy modulation agent can be used alone or as aseries of two or more energy modulation agents wherein the energymodulation agents provide an energy cascade. Thus, the first energymodulation agent in the cascade will absorb the activation energy,convert it to a different energy which is then absorbed by the secondenergy modulation in the cascade, and so forth until the end of thecascade is reached with the final energy modulation agent in the cascadeemitting the energy necessary to activate the photo-activatable agent inthe medium.

Although the photo-activatable agent and the energy modulation agent canbe distinct and separate, it will be understood that the two agents neednot be independent and separate entities. In fact, the two agents may beassociated with each other via a number of different configurations.Where the two agents are independent and separately movable from eachother, they generally interact with each other via diffusion and chanceencounters within a common surrounding medium. Where thephoto-activatable agent and the energy modulation agent are notseparate, they may be combined into one single entity.

The initiation energy source can be any energy source capable ofproviding energy at a level sufficient to activate the activatable agentdirectly, or to provide the energy modulation agent with the inputneeded to emit the activation energy for the activatable agent (indirectactivation). Preferable initiation energy sources include, but are notlimited to, UV-A lamps or fiber optic lines, a light needle, anendoscope, and a linear accelerator that generates x-ray, gamma-ray, orelectron beams. The energy used can be any type, including but notlimited to, gamma ray, x-ray, UV, near-UV, visible, Near IR, IR,microwave, radio wave, etc. In a preferred embodiment, the initiationenergy capable of penetrating completely through the subject. Exemplaryinitiation energy sources that are capable of penetrating completelythrough the subject include, but are not limited to, x-rays, gamma rays,electron beams, microwaves and radio waves.

Basic Principle of Plasmonics and Enhanced Electromagnetic Fields

The plasmonics-enhanced principle is based in theory on enhancementmechanisms of the electromagnetic field effect. These theories areadvanced here for the sake of illustrating the invention and are notnecessarily intended to limit any of the embodiments to this particulartheory. There are two main sources of electromagnetic enhancement: (1)first, the laser electromagnetic field is enhanced due to an addition ofa field caused by a polarization of a metal particle; (2) an enhancementdue to the molecule radiating an amplified emission (luminescence,Raman, etc.) field, which further polarizes the metal particle, therebyacting as an antenna to further amplify a Raman/Luminescence signal.

Electromagnetic enhancements are divided into two main classes: a)enhancements that occur only in the presence of a radiation field, andb) enhancements that occur even without a radiation field. The firstclass of enhancements is further divided into several processes. Plasmaresonances on substrate surfaces, also called surface plasmons, providea significant contribution to electromagnetic enhancement. One effectivetype of plasmonics-active substrate includes nanostructured metalparticles, protrusions, or rough surfaces of metallic materials.Incident light irradiating these surfaces excites conduction electronsin the metal, and induces the excitation of surface plasmons leading toRaman/luminescence enhancement. At a plasmon frequency, metalnanoparticles (or other nanostructured roughened structures) becomepolarized, resulting in large field-induced polarizations and thus largelocal fields on the surface. These local fields increase theluminescence/Raman emission intensity, which is proportional to thesquare of the applied field at the molecule.

As a result, the effective electromagnetic field experienced by ananalyte molecule on these surfaces is much larger than the actualapplied field. This field decreases as 1/r³ away from the surface.Therefore, in the electromagnetic models, the luminescence/Raman-activeanalyte molecule is not required to be in contact with the metallicsurface but can be located anywhere within the range of the enhancedlocal field, which can polarize this molecule. The dipole oscillating atthe wavelength λ of Raman or luminescence can, in turn, polarize themetallic nanostructures and, if λ is in resonance with the localizedsurface plasmons, the nanostructures can enhance the observed emissionlight (Raman or luminescence).

Accordingly, plasmonics-active metal nanoparticles also exhibit stronglyenhanced visible and near-infrared light absorption, several orders ofmagnitude more intense compared to conventional laser phototherapyagents. The use of plasmonic nanoparticles as highly enhancedphotoabsorbing agents thus provides a selective and efficient strategyfor the efficient use of internally generated light.

Accordingly, the invention utilizes several important mechanisms:

-   -   (A) Increased absorption of the excitation light by the        plasmonic metal nanoparticles, resulting in enhanced        photoactivation of photoinitiators or photocatalysts;    -   (B) Increased absorption of the excitation light by the        plasmonic metal nanoparticles that serve as more efficient        energy modulation agent systems, yielding more light for        increased excitation of the photoinitiators or photocatalysts;    -   (C) Increased absorption of the excitation light by the medium        material on or near the plasmonic metal nanoparticles;    -   (D) Increased light absorption of the energy modulation agent        molecules adsorbed on or near the metal nanoparticles;    -   (E) Amplified light emission from the energy modulation agent        molecules adsorbed on or near the metal nanoparticles; and    -   (F) Increased absorption of emission light emitted from the        energy modulation agent by the photoinitiators or        photocatalysts.

As discussed above, one of several phenomena that can enhance theefficiency of light emitted (Raman or luminescence) from moleculesadsorbed or near a metal nanostructures Raman scatter is thesurface-enhanced Raman scattering (SERS) effect. In 1984, the generalapplicability of SERS as an analytical technique was first reported byone of the present inventors, and the possibility of SERS measurementfor a variety of chemicals including several homocyclic and heterocyclicpolyaromatic compounds [T. Vo-Dinh, M. Y.K. Hiromoto, G. M. Begun and R.L. Moody, “Surface-enhanced Raman spectroscopy for trace organicanalysis,” Anal. Chem., vol. 56, 1667, 198-1], the entire contents ofwhich are incorporated herein by reference. Extensive research has beendevoted to understanding and modeling the Raman enhancement in SERSsince the mid 1980's. FIG. 5, for example, shows the early work byKerker modeling electromagnetic field enhancements for spherical silvernanoparticles and metallic nanoshells around dielectric cores as farback as 1984 [M. M. Kerker, Acc. Chem. Res., 17, 370 (1984)], the entirecontents of which are incorporated herein by reference. This figureshows the result of theoretical calculations of electromagneticenhancements for isolated spherical nanospheres and nanoshells atdifferent excitation wavelengths. The intensity of the normally weakRaman scattering process is increased by factors as large as 10¹³ or10¹⁵ for compounds adsorbed onto a SERS substrate, allowing forsingle-molecule detection. As a result of the electromagnetic fieldenhancements produced near nanostructured metal surfaces, nanoparticleshave found increased use as fluorescence and Raman nanoprobes.

Theoretical models indicate that it is possible to tune the size of thenanoparticles and the nanoshells to the excitation wavelength.Experimental evidence suggests that the origin of the 10⁶- to 10¹⁵-foldRaman enhancement primarily arises from two mechanisms: a) anelectromagnetic “lightning rod” effect occurring near metal surfacestructures associated with large local fields caused by electromagneticresonances, often referred to as “surface plasmons,” and b) an effectassociated with direct energy transfer between the molecule and themetal surface.

According to classical electromagnetic theory, electromagnetic fieldscan be locally amplified when light is incident on metal nanostructures.These field enhancements can be quite large (typically 10⁶- to 10⁷-fold,but up to 10¹⁵-fold enhancement at “hot spots”). When a nanostructuredmetallic surface is irradiated by an electromagnetic field (e.g., alaser beam), electrons within the conduction band begin to oscillate ata frequency equal to that of the incident light. These oscillatingelectrons, called “surface plasmons,” produce a secondary electric fieldwhich adds to the incident field. If these oscillating electrons arespatially confined, as is the case for isolated metallic nanospheres orroughened metallic surfaces nanostructures), there is a characteristicfrequency (the plasmon frequency) at which there is a resonant responseof the collective oscillations to the incident field. This conditionyields intense localized field enhancements that can interact withmolecules on or near the metal surface . In an effect analogous to a“lightning rod,” secondary fields are typically most concentrated atpoints of high curvature on the roughened metal surface.

Design, Fabrication and Operation of Plasmonics-enhanced Structures

FIGS. 6A-6G shows a number of the various embodiments ofplasmonics-enhanced probe structures (PEPST) that can be designed:

-   -   (A) Photo-activatable (PA) molecules bound to a metal (e.g.,        gold) nanoparticle;    -   (B) Photo-activatable (PA) molecule covered with metal        nanoparticles;    -   (C) Metal nanoparticle covered with PA nanocap;    -   (D) PA-containing nanoparticle covered with metal nanocap;    -   (E) Metal nanoparticle covered with PA nanoshell;    -   (F) PA-containing nanoparticle covered with metal nanoshell; and    -   (G) PA-containing nanoparticle covered with metal nanoshell with        protective coating layer.

A basic embodiment of the PEPST is shown in FIG. 6A. This PEPST includesPA molecules bound to a metal (e.g., gold) nanoparticle. FIG. 7illustrates the plasmonics-enhancement effect as it would be used inthis invention to enhance the interaction of the primary excitationlight source with energy modulation agents or to enhance the interactionof the secondarily produced light with the medium in effecting a changeto the medium. Radiation of suitable energy is used to excite the PEPSTstructures which in turn activates for example nearby photoinitiators.

For example, light of a HeNe laser (632.8-nm excitation) can be used forexcitation. In this case the metal nanoparticles are designed to exhibitstrong plasmon resonance band around 632.8 nm. The surface plasmonresonance effect amplifies the excitation light at the nanoparticles,resulting in an increased photoactivation of a photo-initiator or aphoto-catalyst and improved reaction kinetic. Further, for sterilizationapplications, the effect increases the likelihood for a germicide eventin the medium in vicinity of the nanoparticles. While light such as theHeNe laser light might be scattered and absorbed in the medium, thepresence of the PEPST structures enhances the interaction of thepenetrating light beyond that which would normally be considered useful.The plasmonics-enhanced mechanism can also be used with the other PEPSTprobes in FIGS. 6B, 6C, 6D, 6E, 6F and 6G.

Structures of Plasmonics-Active Metal Nanostructures

Plasmon resonances arise within a metallic nanoparticle from thecollective oscillation of free electrons driven by an incident opticalfield. The plasmon c response of nanoparticles have played a role in agrowing number of applications, including surface-enhanced Ramanscattering (SERS), chemical sensing, drug delivery, photothermal cancertherapy, and new photonic devices. The investigation and application ofplasmonics nanosubstrates for SERS detection has been used by one of thepresent inventors for over two decades [T. Vo-Dinh, “Surface-EnhancedRaman Spectroscopy Using Metallic Nanostructures,” Trends in Anal.Chem., 17, 557 (1998)], the entire contents of which are incorporatedherein by reference. The first report by one of the present inventors onthe practical analytical use of the SERS techniques for trace analysisof a variety of chemicals including several homocyclic and heterocyclicpolyaromatic compounds was in 1984 [T. Vo-Dinh, M. Y. K. Hiromoto, G. M.Begun and R. L. Moody, “Surface-enhanced Raman spectroscopy for traceorganic analysis,” Anal. Chem., vol. 56, 1667, 1984], the entirecontents of which are incorporated herein by reference. Since then, thedevelopment of SERS technologies for applications in chemical sensing,biological analysis and medical diagnostics has been ongoing. Thesubstrates involve nanoparticles and semi-nanoshells having a layer ofnanoparticles coated by a metal (such as silver) on one side (nanocapsor half-shells). Several groups have shown that plasmon resonances ofspherical shells can be tuned by controlling the shell thickness andaspect ratios of the nanoshell structures [M. M. Kerker, Acc. Chem.Res., 17, 370 (1984); J. B. Jackson, S. L. Westcott, L. R. Hirsch, J. L.West and N. H. Halas, “Controlling the surface enhanced Raman effect viathe nanoshell geometry,” Appl. Phys. Lett., vol. 82, 257-259, 2003, theentire contents of which are incorporated herein by reference; S. J.Norton and T Vo-Dinh, “Plasmonic Resonances of nanoshells of SpheroidalShape”, IEEE Trans. Nanotechnology, 6, 627-638 (2007) , the entirecontents of which are incorporated herein by reference ]. These shellstypically have a metallic layer over a dielectric core. In oneembodiment of the invention, these shells include spheroidal shells,since the plasmon resonances (both longitudinal and transverse modes)are influenced by both shell thickness and aspect ratio. A number ofresearchers have examined the plasmonic response of the solid spheroidalparticle in their analysis of surface-enhanced Raman scattering,although the spheroidal shell appears not to have been investigated. Theinvention also includes prolate and oblate spheroidal shells, which showsome interesting qualitative features in their plasmon resonances. Thespheroidal shell presents two degrees of freedom for tuning: the shellthickness and the shell aspect ratio [S. J. Norton and T. Vo-Dinh,“Plasmonic Resonances of Nanoshells of Spheroidal Shape”, IEEE Trans.Nanotech ology, 6, 627-638 (2007)], the entire contents of which areincorporated herein by reference.

FIG. 7 shows some of the various embodiments of plasmonics-activenanostructures that can be designed, and are preferred embodiments ofthis invention:

-   -   (A) Metal nanoparticle;    -   (B) Dielectric nanoparticle core covered with metal nanocap;    -   (C) Spherical metal nanoshell covering dielectric spheroid core;    -   (D) Oblate metal nanoshell covering dielectric spheroid core;    -   (E) Metal nanoparticle core covered with dielectric nanoshell;    -   (F) Metal nanoshell with protective coating layer;    -   (G) Multi layer metal nanoshells covering dielectric spheroid        core;    -   (H) Multi-nanoparticle structures;    -   (I) Metal nanocube and nanotriangle/nanoprism; and    -   (J) Metal cylinder.        PEPST Probes with Remotely-Activated Photoactivatable Molecules

In a further embodiment of the invention, the PA molecules can beincorporated into a material (e.g., biocompatible polymer) that can forma nanocap onto the metal (gold) nanoparticles. The material can be a gelor biocompatible polymer that can have long-term continuous releaseproperties. Suitable gel or biocompatible polymers include, but are notlimited to poly(esters) based on polylactide (PLA), polyglycolide (PGA),polycarpolactone (PCL), and their copolymers, as well aspoly(hydroxyalkanoate)s of the PHB-PHV class, additional poly(ester)s,natural polymers, particularly, modified poly(saccharide)s, e.g.,starch, cellulose, and chitosan, polyethylene oxides, poly(ether)(ester)block copolymers, and ethylene vinyl acetate copolymers. The releasemechanism can also be triggered by non-invasive techniques, such as RF,MW, ultrasound, photon (FIG. 8).

FIG. 9 shows other possible embodiments where the PA molecule is boundto the metal nanoparticles via a linker that can be cut by a photonradiation. Such a linker includes, but is not limited to, a biochemicalbond (FIG. 9A), a DNA bond (FIG. 9B), or an antibody-antigen bond (FIG.9C). In another embodiment, the linker is a chemically labile bond thatwill be broken by the chemical environment inside the cell. In variousembodiments, it may be more difficult for metal nanoparticles to entertargeted cites in the medium than for smaller molecules. In theseembodiments, it is desirable to have PEPST probes that have releasablePA molecules.

Aggregation of metal (such as silver or gold) nanoparticles(nanospheres, nanorods, etc) is often a problem, especially withcitrate-capped gold nanospheres, cetyl trimethylammonium bromide(CTAB)-capped gold nanospheres and nanorods and nanoshells because theyhave poor stability when they are dispersed in buffer solution due tothe aggregating effect of salt ions. The biocompatibility can beimproved and nanoparticle aggregation prevented by capping thenanoparticles with polyethylene glycol (PEG) (by conjugation ofthiol-functionalized PEG with metal nanoparticles).

Immobilization of Biomolecules and Metal Nanoparticles

The immobilization of biomolecules (PA molecules, drugs, proteins,enzymes, antibodies, DNA, etc.) to a support can use a wide variety ofmethods published in the literature. For example, the encapsulatedstructures 10 of FIG. 3C and 3D can be modified in one embodiment ofthis invention such that the PEPST structures are immobilized on theouter exposed surfaces such that any light from the encapsulatedstructures would be enhanced in interaction with the medium.Furthermore, in one embodiment, the encapsulated structures 10 can notinclude an energy modulation agent. Rather, light from an externalsource such as a flash lamp or a LED array or laser or UV source couldbe transmitted through the empty encapsulated structures 10 andpropagate into the medium. Binding can be performed through covalentbonds taking advantage of reactive groups such as amine (—NH₂) orsulfide (—SH) that naturally are present or can be incorporated into thebiomolecule structure. Amines can react with carboxylic acid or estermoieties in high yield to form stable amide bonds. Thiols canparticipate in maleimide coupling, yielding stable dialkylsulfides.

One solid support of interest in this invention is the metal (preferablygold or silver) nanoparticles. The majority of immobilization schemesinvolving metal surfaces, such as gold or silver, utilize a priorderivatization of the surface with alkylthiols, forming stable linkages.Alkylthiols readily form self-assembled monolayers (SAM) onto silversurfaces in micromolar concentrations. The terminus of the alkylthiolchain can be used to bind biomolecules, or can be easily modified to doso. The length of the alkylthiol chain has been found to be an importantparameter, keeping the biomolecules away from the surface, with lengthsof the alkyl group from 4 to 20 carbons being preferred.

There are many methods related to the preparation of stableoligonucleotide conjugates with gold particles by usingthiol-functionalized biomolecules that had previously been shown to formstrong gold-thiol bonds. Oligonucleotides with 5′-terminal alkanethiolfunctional groups as anchors can be bound to the surface of goldnanoparticles, and the resulting labels were robust and stable to bothhigh and low temperature conditions [R. Elghanian, J. J. Storhoff, R. C.Mucic, R. L. Letsinger and C. A. Mirkin, Selective colorimetricdetection of polynucleotides based on the distance-dependent opticalproperties of gold nanoparticles. Science 277 (1997), pp. 1078-1081],the entire contents of which are incorporated herein by reference. Acyclic dithiane-epiandrosterone disulfide linker has been developed forbinding oligonucleotides to gold surfaces [R. Elghanian, J. J. StorhoffR. C. Mucic, R. L. Letsinger and C. A. Mirkin, Selective colorimetricdetection of polynucleotides based on the distance-dependent opticalproperties of gold nanoparticles. Science 277 (1997), pp. 1078-1081],the entire contents of which are incorporated herein by reference. Li etal. have reported a trithiol-capped oligonucleotide that can stabilizegold metal nanoparticles having diameters=100 nm, while retaininghybridization properties that are comparable to acyclic ordithiol-oligonucleotide modified particles [Z. Li, R. C. Jin, C. A.Mirkin and R. L. Letsinger, Multiple thiol-anchor capped DNA-goldnanoparticle conjugates. Nucleic Acids Res. 30 (2002), pp. 1558-1562],the entire contents of which are incorporated herein by reference.

In general silver nanoparticles cannot be effectively passivated byalkylthiol-modified oligonucleotides using the established experimentalprotocols that were developed for gold particles. One method ofgenerating core-shell particles having a core of silver and a thin shellof gold has allowed silver nanoparticles to be readily functionalizedwith alkylthiol-oligonucleotides using the proven methods used toprepare pure gold particle-oligonucleotide conjugates. [Y. W. Cao, R.Jill and C. A. Mirkin, DNA-modified core-shell Ag/Au nanoparticles. J.Am. Chem. Soc. 123 (2001), pp. 7961-7962], the entire contents of whichare incorporated herein by reference.

Silver surfaces have been found to exhibit controlled self-assemblykinetics when exposed to dilute ethanolic solutions of alkylthiols. Thetilt angle formed between the surface and the hydrocarbon tail rangesfrom 0 to 15°. There is also a larger thiol packing density on silver,when compared to gold [Burges, J. D.; Hawkridge, F. M. Langmuir 1997,13, 3781-6], the entire contents of which are incorporated herein byreference. After self-assembled monolayer (SAM) formation on gold/silvernanoparticles, alkylthiols can be covalently coupled to biomolecules.The majority of synthetic techniques for the covalent immobilization ofbiomolecules utilize free amine groups of a polypeptide (enzymes,antibodies, antigens, etc) or of amino-labeled DNA strands, to reactwith a carboxylic acid moiety forming amide bonds.

Such bonding schemes have applications not only by providing a mechanismby which the nanoparticles can be controllably dispersed and deliveredwithin a medium, but may also play a role in the formation of theencapsulated structures of the invention, as detailed above.

Spectral Range of Light Used for PEPST

A plasmonics enhanced effect can occur throughout the electromagneticregion provided the suitable nanostructures, nanoscale dimensions, metaltypes are used. Therefore, the PEPST concept can be utilized for theentire electromagnetic spectrum, i.e, energy, ranging from gamma raysand X rays throughout ultraviolet, visible, infrared, microwave andradio frequency energy. However, for practical reasons, visible and NIRlight are used for silver and gold nanoparticles, since the plasmonresonances for silver and gold occur in the visible and NIR region,respectively. Especially for gold nanoparticles, the NIR region is veryappropriate for the delivery of energy into a medium where otherwiseoptical scatter at shorter wavelengths would present a problem, such asfor example in the treatment of waste water or the sterilization of foodproducts having high concentrations of suspended solids.

Photon Excitation

There are several methods of the invention for using light to excitephotoactivate compounds in the medium. One can use light havingwavelengths within the so-called “window” (designed to penetrate anycontainer holding the medium to be processed and/or to transmit hroughthe medium). Moreover, while certain aspects of the invention preferthat the excitation light be nominally non-absorbing in the medium, dueto the plasmonic advantages, the invention is still useful in mediumswhere there is considerable scatter and absorption. For example, in theabove-noted, UV applications, the plasmonic enhanced PEPST probes couldbe introduced into the medium and UV light could be used as theactivation source. While in the region of the medium near the surface,the PEP ST probes may not play a dominant role, in regions deeper intothe surface where the UV light has become attenuated, the PEPST probeswill play a significant role in photoinitiation or photo-catalyst.

The ability of light to penetrate the medium depends on absorption andscatter. Within the hydrous medium, a window extends from 600 to 1300nm, from the orange/red region of the visible spectrum into the NIR. Atthe short-wavelength end, absorbing biomolecules become important,including DNA and the amino acids tryptophan and tyrosine. At theinfrared (TR) end of the window, penetration is limited by theabsorption properties of water. Within the window, scattering isdominant over absorption, and so the propagating light becomes diffuse,although not necessarily entering into the diffusion limit. FIG. 10shows a diagram of the window for an hydrous medium. The followingsection discusses the use of one-photon and multi-photon techniques.

Light Excitation Methods: Single-Photon and Multi-Photon Excitation

Two methods can be used, one-photon or multi-photon excitation. If thetwo-photon technique is used, one can excite the PA molecules with lightat 700-1000 nm, which can penetrate deep inside the medium, in order toexcite molecules that absorb in the 350-500 nm spectral region. Thisapproach can excite compounds, which absorb in the 290-350 nm spectralregion and emit in the visible. With the one-photon method, thephoto-activator (PA) molecules can directly absorb excitation light at600-1300 nm. In this case we can design a system having additionalaromatic rings or other conjugation to alter the ability to absorb atdifferent wavelengths.

X Ray Excitation

Although X-ray can excite compounds in a medium non-invasively, X-ray isnot easily absorbed by many of the compounds where energy modulation isdesired. This invention provides a solution to this problem, by theproviding of a molecular system that can absorb the X-ray energy andchange that energy into other energies that can be used. Morespecifically, one example of a molecular system that can absorb andchange the X-ray energy in this invention is the PEPST probes includingnanoparticles (as described above).

In this embodiment, the invention uses X-rays for excitation. Theadvantage is the ability to excite molecules non-invasively since X-raycan penetrate deep in the medium. In one embodiment of the invention, aPA molecule (e.g., a photoinitiator) is bound to a molecular entity,referred to as an “energy modulation agent” that can interact with theX-rays, and then the emitted light that can be absorbed by the PAmolecules. (FIG. 11)

PEPST Probes for X Ray Excitation

In the previous sections, the advantage of gold nanoparticles asplasmonics-active systems have been discussed. Furthermore, goldnanoparticles are also suitable energy modulation agent systems sincethey are biocompatible and have been shown to be a possible candidatefor contrast agents for X-ray [Hainfeld et al, The British Journal ofradiology, 79, 248, 2006], the entire contents of which are incorporatedherein by reference. The concept of using high-Z materials for doseenhancement in cancer radiotherapy was advanced over 20 years ago. Theuse of gold nanoparticles as a dose enhancer seems more promising thanthe earlier attempts using microspheres and other materials for twoprimary reasons. First, gold has a higher Z number than iodine (I, Z=53)or gadolinium (Gd, Z=64), while showing little toxicity, up to at least3% by weight, on either the rodent or human tumour cells. The goldnanoparticles were non-toxic to mice and were largely cleared from thebody through the kidneys. This novel use of small gold nanoparticlespermits material which may incidentally uptake some of thesenanoparticles to remain safe for human consumption.

FIG. 12 shows a number of the various embodiments of PEPST probes thatcan be preferably used for X ray excitation of energy modulationagent-PA system. These probes comprise:

-   -   (A) PA molecules bound to energy modulation agent and to        plasmonic metal nanoparticle;    -   (B) Plasmonic metal nanoparticle with energy modulation agent        nanocap covered with PA molecules;    -   (C) PA-covered nanoparticle with plasmonic metal nanoparticles;    -   (D) Energy modulation agent-containing nanoparticle covered with        PA molecules and plasmonic metal nanocap;    -   (E) Plasmonic metal nanoparticle core with energy modulation        agent nanoshell covered with PA molecule; and    -   (F) PA molecule bound to energy modulation agent (attached to        plasmonics metal nanoparticle) nanoparticle by detachable        biochemical bond.

Examples of PEPST System Based on Energy Modulation Agent-PA

For purposes of simplification, the following discussion is centered ongold as the metal material and CdS as the energy modulation agentmaterial (which can also be used as DNA stabilized CdS, see Ma et al,Langmuir, 23 (26), 12783-12787 (2007), the entire contents of which areincorporated herein by reference). However, it is to be understood thatmany other embodiments of metal material, energy modulation agent and PAmolecule are possible within the bounds of the invention, and thefollowing discussion is for exemplary purposes only.

In the embodiment of FIG. 12A, the PEPST system comprises goldnanoparticles, an energy modulation agent nanoparticle (e.g., CdS)linked to a PA drug molecule (e.g., psoralen). X ray is irradiated toCdS, which absorbs X rays [Hua et al, Rev. Sci. Instrum. ,, 73, 1379,2002, the entire contents of which are incorporated herein by reference]and emits CdS XEOL light (at 350-400 nm) that is plasmonics-enhanced bythe gold nanoparticle. This enhanced XEOL light can be used tophotoactivate PA molecules. In this case the nanostructure of the goldnanoparticle is designed to enhance the XEOL light at 350-400 nm.

In the embodiment of FIG. 12B, the PEPST system comprises aplasmonics-active metal (gold) nanoparticle with energy modulation agentnanocap (CdS) covered with PA molecules. X ray is irradiated to CdS,which absorbs X ray and emits XEOL light that is plasmonics-enhanced bythe gold nanoparticle. This enhanced XEOL light is used to photoactivatethe PA molecule.

In the embodiment of FIG. 12C, the PEPST system comprises a PA (e.g.,psoralen covered CdS nanoparticle with smaller plasmonic metal (gold)nanoparticles. X ray is irradiated to CdS, which absorbs X ray and emitsXEOL light that is plasmonics-enhanced by the gold nanoparticle. Thisenhanced XEOL light is used to photoactivate the PA molecule.

In the embodiment of FIG. 12D, the energy modulation agent corecomprises CdS or CsCl nanoparticles covered with a nanocap of gold. Xray is irradiated to CdS or CsCl, which absorbs X ray [[Jaegle et al, J.Appl. Phys., 81, 2406, 1997] and emits XEOL light that isplasmonics-enhanced by the gold nanocap structure. This enhanced XEOLlight is used to photoactivate the PA molecule.

Similarly, the embodiment in FIG. 12E comprises a spherical gold corecovered by a shell of CdS or CsCl. X ray is irradiated to CdS or CsClmaterial, which absorbs X ray [Jaegle et al, J. Appl. Phys., 81, 2406,1997, the entire contents of which are incorporated herein by reference]and emits XEOL light that is plasmonics-enhanced by the gold nanosphere.This enhanced XEOL light is used to photoactivate the PA molecule.

In the embodiment of FIG. 12F, the PEPST system comprises goldnanoparticles, and an energy modulation agent nanoparticle (e.g., CdS)linked to a PA drug molecule (e.g., psoralen) by a link that can bedetached by radiation. X ray is irradiated to CdS, which absorbs X rayand emits CdS XEOL light (at 350-400 nm) that is plasmonics-enhanced bythe gold nanoparticle. This enhanced XEOL light is used to photoactivatepsoralen (PA molecule). In this case the nanostnicture of the goldnanoparticle is designed to enhance the XEOL light at 350-400 nm.

In alternative embodiments, the metal nanoparticles or single nanoshellsare replaced by multi layers of nanoshells [Kun Chen, Yang Liu,Guillermo Ameer, Vadim Backman, Optimal design of structured nanospheresfor ultrasharp light-scattering resonances as molecular imagingmultilabels, Journal of Biomedical Optics, 10(2), 024005 (March/April2005), the entire contents of which are incorporated herein byreference].

In other alternative embodiments the metal nanoparticles are coveredwith a layer (1-30 nm) of dielectric material (e.g. silica). Thedielectric layer (or nanoshell) is designed to prevent quenching of theluminescence light emitted by the energy modulation agent (also referredto as EEC) molecule(s) due to direct contact of the metal with theenergy modulation agent molecules. In yet other alternative embodiments,the energy modulation agent molecules or materials are bound to (or inproximity of) a metal nanoparticle via a spacer (linker). The spacer isdesigned to prevent quenching of the luminescence light emitted by theenergy modulation agent molecules or materials.

Other Useable Materials

The energy modulation agent materials can include any materials that canabsorb X ray and emit light in order to excite the PA molecule. Theenergy modulation agent materials include, but are not limited to:

metals (gold, silver, etc);

quantum dots;

semiconductor materials;

scintillation and phosphor materials;

materials that exhibit X-ray excited luminescence (XEOL);

organic solids, metal complexes, inorganic solids, crystals, rare earthmaterials (lanthanides), polymers, scintillators, phosphor materials,etc.; and

materials that exhibit excitonic properties.

Quantum dots, semiconductor nanostructures. Various materials related toquantum dots, semiconductor materials, etc. can be used as energymodulation agent systems. For example CdS-related nanostructures havebeen shown to exhibit X-ray excited luminescence in the UV-visibleregion [Hua et al, Rev. Sci. Instrum., 73, 1379, 2002, the entirecontents of which are incorporated herein by reference].

Scintillator materials as energy modulation agent systems. Variousscintillator materials can be used as energy modulation agents sincethey absorb X-ray and emit luminescence emission, which can be used toexcite the PA system. For example, single crystals of molybdates can beexcited by X-ray and emit luminescence around 400 nm [Mirkhin et al,Nuclear Instrum. Meth. In Physics Res. A, 486, 295 (2002, the entirecontents of which are incorporated herein by reference].

Solid materials as energy modulation agent systems: Various solidmaterials can be used as energy modulation agents due to their X-rayexcited luminescence properties. For example CdS (or CsCl) exhibitluminescence when excited by soft X-ray [Jaegle et al, J. Appl. Phys.,81, 2406, 1997, the entire contents of which are incorporated herein byreference].

XEOL materials: lanthanides or rare earth materials; see L. Soderholm,G. K. Liu, Mark R. Antonioc, F. W. Lytle, X-ray excited opticalluminescence .XEOL. detection of x-ray absorption fine structure .XAFZ,J. Chem. Phys,109, 6745, 1998, the entire contents of which areincorporated herein by reference or, Masashi Ishiia, Yoshihito Tanakaand Tetsuya Ishikawa, Shuji Komuro and Takitaro Morikawa, YoshinobuAoyagi, Site-selective x-ray absorption fine structure analysis of anoptically active center in Er-doped semiconductor thin film usingx-ray-excited optical luminescence, Appl. Phys. Lett, 78, 183, 200, theentire contents of which are incorporated herein by reference.

Some examples of metal complexes exhibiting XEOL which can be used asenergy modulation agent systems are shown in FIGS. 13 and 14. Suchstructures can be modified by replacing the metal atom with metalnanoparticles in order to fabricate a plasmonics-enhance PEPSI probe. Inthe invention, the experimental parameters including size, shape andmetal type of the nano structure can be selected based upon theexcitation radiation (NIR or X ray excitation), the photoactivationradiation (UVB), and/or the emission process from the energy modulationagent system (visible NIR)

Principle of Plasmonics-Enhancement Effect of the PEPST Probe UsingX-Ray Excitation

One embodiment of the basic PEPSI probe embodiment comprises PAmolecules bound to an energy modulation agent and to plasmonic metal(gold) nanoparticles. The metal nanoparticle can play 2 roles:

-   -   (A) Enhancement of the X-ray electromagnetic field    -   (B) Enhancement of the emission signal of the energy modulation        agent system.

The X ray radiation, used to excite the energy modulation agent system,is amplified by the metal nanoparticle due to plasmon resonance. As aresult the energy modulation agent system exhibits more emission lightthat s used to photoactivate the PA molecules and make them photoactive.In this case the metal nanoparticles are designed to exhibit strongplasmon resonance at or near the X ray wavelengths. The surface plasmonresonance effect amplifies the excitation light at the nanoparticles,resulting in increased photoactivation of the PA drug molecules andimproved therapy efficiency. The plasmonics-enhanced mechanism can alsobe used with the other PEPSI probes described above.

FIG. 15 illustrates the plasmonics-enhancement effect of the PEPSIprobe. X-ray used in medical diagnostic imaging has photon energies fromapproximately 10 to 150 keV, which is equivalent to wavelengths rangefrom 1.2 to 0.0083 Angstroms. [λ (Angstrom)=12.4/E (keV)]. Soft X raycan go to 10 nm. The dimension of plasmonics-active nanoparticlesusually have dimensions on the order or less than the wavelengths of theradiation used. Note that the approximate atomic radius of gold isapproximately 0.15 nanometers. At the limit, for gold the smallest“nanoparticle” size is 0.14 nm (only 1 gold atom). A nanoparticle withsize in the hundreds of nm will have approximately 10⁶-10⁷ gold atoms.Therefore, the range of gold nanoparticles discussed in this inventioncan range from 1- 10⁷gold atoms.

The gold nanoparticles can also enhance the energy modulation agentemission signal, which is use to excite the PA molecule. For psoralens,this spectral range is in the UVB region (320-400nm). Silver or goldnanoparticles, nanoshell and nanocaps have been fabricated to exhibitstrong plasmon resonance in this region. FIG. 16 shows excitation andemission fluorescence spectra of a psoralen compound (8-methoxypsoralen)

Nanoparticle Chain for Dual Plasmonics Effect

As discussed previously, there is the need to develop nanoparticlesystems that can have dual (or multi) plasmonics resonance modes. FIG.17 illustrates an embodiment of the invention PEPST probe having a chainof metal particles having different sizes and coupled to each other,which could exhibit such dual plasmonics-based enhancement. For examplethe parameters (size, metal type, structure, etc) of the largernanoparticle (FIG. 17, left) can be tuned to NIR, VIS or UV light whilethe smaller particle (FIG. 17, right) can be tuned to X ray. There isalso a coupling effect between these particles.

These nanoparticle chains are useful in providing plasmonics enhancementof both the incident radiation used (for example, x-ray activation ofCdS) as well as plasmonics enhancement of the emitted radiation thatwill then activate the PA. Similar nanoparticles systems have been usedas nanolens [Self-Similar Chain of Metal Nanospheres as an EfficientNanolens, Kuiru Li, Mark I. Stockman, and David J. Bergman, PhysicalReview Letter, VOLUME 91, NUMBER 22, 227402-1, 2003, the entire contentsof which are incorporated herein by reference].

Fabrication of Gold Nanoparticles: The Frens method [Frens, G.,Controlled nucleation for the regulation of the particle size inmonodisperse gold solutions. Nature (London) Phys Sci,, 1973. 241: p.20-22, the entire contents of which are incorporated herein byreference] can be used in the invention to synthesize a solution of goldnanoparticles ranging in diameter from 8-10 nm. Briefly, 5.0×10⁻⁶ mol ofHAuCl₄ is dissolved in 19 ml of deionized water producing a faintyellowish solution. This solution is heated with vigorous stirring in arotary evaporator for 45 minutes. 1 ml of 0.5% sodium citrate solutionis added and the solution is stirred for an additional 30 minutes. Thecolor of the solution gradually changed from the initial faint yellowishto clear, grey, purple and finally a tantalizing wine-red color similarto merlot. The sodium citrate used serves in a dual capacity, firstacting as a reducing agent, and second, producing negative citrate ionsthat are adsorbed onto the gold nanoparticles introducing surface chargethat repels the particles and preventing nanocluster formation.

Another method for synthesizing gold nanoparticles involvesstabilization by horse spleen apoferritin (HSAF) has been reported usingNaBH₄ or 3-(N-morpholino) propanesulfonic acid (MOPS) as the reducingagent [Lei Zhang, Joe Swift, Christopher A. Butts, Vijay Yerubandi andIvan J. Dmochowski, Structure and activity of apoferritin-stabilizedgold nanoparticles, Journal of Inorganic Biochemistry, Vol. 101,1719-1729, 2007, the entire contents of which are incorporated herein byreference]. Gold sulfite (Au₂S) nanoparticles were prepared in thecavity of the cage-shaped protein, apoferritin. Apoferritin has acavity, 7 nm in diameter, and the diameter of fabricated Au₂Snanoparticles is about the same size with the cavity and size dispersionwas small. [Keiko Yoshizawa, Kenji Iwahori, Kenji Sugimoto and IchiroYamashita, Fabrication of Gold Sulfide Nanoparticles Using the ProteinCage of Apoferritin, Chemistry Letters, Vol. 35 (2006) , No. 10 p.1192,the entire contents of which are incorporated herein by reference].Thus, in one embodiment, the PA or energy modulation agent-PA compoundsare encapsulated inside the apoferrtin shells.

Excitons in Solid Materials

Excitons are often defined as “quasiparticles” inside a solid material.In solid materials, such as semiconductors, molecular crystals andconjugated organic materials, light excitation at suitable wavelength(such as X ray, UV and visible radiation, etc) can excite electrons fromthe valence band to the conduction band. Through the Coulombinteraction, this newly formed conduction electron is attracted, to thepositively charged hole it left behind in the valence band. As a result,the electron and hole together form a bound state called an exciton.(Note that this neutral bound complex is a “quasiparticle” that canbehave as a boson—a particle with integer spin which obeys Bose-Einsteinstatistics; when the temperature of a boson gas drops below a certainvalue, a large number of bosons ‘condense’ into a single quantumstate—this is a Bose-Einstein condensate (BEC). Exciton production isinvolved in X-ray excitation of a solid material. Wide band-gapmaterials are often employed for transformination of the x-ray toultraviole/uvisible photons in the fabrication of scintillators andphosphors [Marlin Nikl, Scintillation dectectors for x-rays, Meas. Sci.Technol. 17 (2006) R37-R54 the entire contents of which are incorporatedherein by reference]. The theory of excitons is well known in materialsresearch and in the fabrication and applications of semiconductors andother materials.

During the initial conversion a multi-step interaction of a high-energyX-ray photon with the lattice of the scintillator material occursthrough the photoelectric effect and Compton scattering effect; forX-ray excitation below 100 keV photon energy the photoelectric effect isthe main process. Many excitons (i.e., electron-hole pairs) are producedand thermally distributed in the conduction bands (electrons) andvalence bands (holes). This first process occurs within less than 1 ps.In the subsequent transport process, the excitons migrate through thematerial where repeated trapping at defects may occur, leading to energylosses due to nonradiative recombination, etc. The final stage,luminescence, consists in consecutive trapping of the electron-holepairs at the luminescent centers and their radiative recombination. Theelectron-hole pairs can be trapped at the defects and recombine,producing luminescent. Luminescent dopants can also be used as traps forexciton.

Exciton Traps

Exciton traps can be produced using impurities in the crystal hostmatrix. In impure crystals with dipolar guest molecules, electron trapstates may arise when an electron is localized on a neighbor of theimpurity molecule. Such traps have been observed in anthracene dopedwith carbazole [Kadshchuk, A. K., Ostapenko, N. I., Skryshevskii, Yu.A., Sugakov, V. I. and Susokolova, T O., Mol. Cryst. and Liq. Cryst.,201, 167 (1991) the entire contents of which are incorporated herein byreference]. The formation of these traps is due to the interaction ofthe dipole moment of the impurity with charge carrier. When theconcentration of the dopant (or impurities) is increased, spectraexhibit additional structure of spectrum due to the trapping of carrierson clusters of impurity molecules. Sometimes, impurities and dopants arenot required: The electron or exciton can also be trapped on astructural defect in such crystals due to the electrostatic interactionwith reoriented dipole moment of disturbed crystal molecules [S. V.Izvekov, V. I. Sugakov, Exciton and Electron Traps on Structural Defectsin Molecular Crystals with Dipolar Molecules, Physica Scripta. Vol. T66,255-257, 1996]. One can design structural defects in molecular crystalsthat serve as exciton traps. The development of GaAs/AlGaAsnanostructures and use of nanofabrication technologies can permitengineered exciton traps with novel quantum mechanical properties inmaterials to be used in the invention.

Design, Fabrication and Operation of EIP Probes

FIG. 18A-D shows various embodiments of EIP probes that can be designed:

-   -   (A) probe comprising PA molecules bound (through a linker, which        can be fixed or detachable) to an energy modulation agent        particle that can produce excitons under radiative excitation at        a suitable wavelength (e.g., X-ray). In this preferred        embodiment, the energy modulation agent materials have        structural defects that serve as traps for excitons.    -   (B) probe comprising PA molecules bound (through a linker, which        can be fixed or detachable) to an energy modulation agent        particle that can produce excitons under radiative excitation at        a suitable wavelength (e.g., X-ray). In this preferred        embodiment, the energy modulation agent materials have        impurities or dopant molecules that serve as traps for excitons.

EIP Probes with Tunable Emission:

The probes as described above in (B) provide the capability to tune theenergy conversion from an X ray excitation source into a wavelength ofinterest to excite the PA molecules. In 1976, D'Silva et al demonstratedthat polynuclear aromatic hydrocarbons (PAH) molecules doped in a frozenn-alkane solids could be excited by X-ray and produce luminescence atvisible wavelengths characteristics of their luminescence spectra. [A.P. D'Silva, G. J. Oestreich, and V. A. Fassel, X-ray excited opticalluminescence of polynuclear aromatic hydrocarbons, Anal. Chem.; 1976;48(6) pp 915-917, the entire contents of which are incorporated hereinby reference]. Tunable EIP probes can be designed to contain suchluminescent dopants such as highly luminescent PAHs exhibitingluminescence emission in the range of 300-400 nm suitable to activatepsoralen. One embodiment of the EIP with tunable emission includes asolid matrix (semiconductors, glass, quartz, conjugated polymers, etc)doped with naphthalene, phenanthrene, pyrene or other compoundsexhibiting luminescence (fluorescence) in the 300-400nm range [T.Vo-Dinh, Multicomponent analysis by synchronous luminescencespectrometry, Anal. Chem.; 1978; 50(3) pp 396-401 the entire contents ofwhich are incorporated herein by reference]. The EEC matrix could be asemiconductor material, preferably transparent at optical wavelength ofinterest (excitation and emission).

Other dopant species such as rare earth materials can also be used asdopants. FIG. 19 shows the X ray excitation optical luminescence (XEOL)of Europium doped in a matrix of BaFBr, emitting at 370-420 nm. U.S.Patent Application Publication No. 2007/0063154 (hereby incorporated byreference) describes these and other nanocomposite materials (andmethods of making them) suitable for XEOL.

FIG. 20 shows various embodiments of EIP probes that can be designed:

(A) probe comprising PA molecules bound around the energy modulationagent particle or embedded in a shell around an energy modulation agentparticle that can produce excitons under radiative excitation at asuitable wavelength (e.g., X-ray). In this embodiment, the energymodulation agent materials have structural defects that serve as trapsfor excitons.

(A) probe comprising PA molecules bound around the energy modulationagent particle or embedded in a shell around an energy modulation agentparticle that can produce excitons under radiative excitation at asuitable wavelength (e.g., X-ray). In this embodiment, the energymodulation agent materials have impurities or dopant molecules thatserve as traps for excitons.

A fundamental key concept in photophysics is the formation of newquasiparticles from admixtures of strongly-coupled states. Such mixedstates can have unusual properties possessed by neither originalparticle. The coupling between excitons and plasmons can be either weakor strong. When the light-matter interaction cannot be considered as aperturbation, the system is in the strong coupling regime. A strongcoupling between a surface plasmon (SP) mode and organic excitons occurshas been shown; the organic semiconductor used was a concentratedcyanine dye in a polymer matrix deposited on a silver film [Ref J.Bellessa, * C. Bonnand, and J. C. Plenet, J. Mugnier, Strong Couplingbetween Surface Plasmons and Excitons in an Organic Semiconductor, Phys.Rev. Lett, 93 (3), 036404-1, 2004, the entire contents of which areincorporated herein by reference]. Other work ahs described thephotophysical properties of excitons in hybrid complexes consisting ofsemiconductor and metal nanoparticles. The interaction betweenindividual nanoparticles can produce an enhancement or suppression ofemission. Enhanced emission comes from electric field amplified by theplasmon resonance, whereas emission suppression is a result of energytransfer from semiconductor to metal nanoparticles. [Alexander 0.Govorov, Garnett W. Bryant,‡ Wei Zhang, Timur Skeini, Jaebeom Lee,§Nicholas A. Kotov, Joseph M. Slocik, and Rajesh R. Naik, Exciton-PlasmonInteraction and Hybrid Excitons in Semiconductor-Metal NanoparticleAssemblies, Nano Lett., Vol. 6, No. 5, 984, 2006, the entire contents ofwhich are incorporated herein by reference]. Other work has described atheory for the interactions between excitonic states and surfaceelectromagnetic modes in small-diameter (<1 nm) semiconductingsingle-walled carbon nanotubes (CNs). [I. V Bondarev, K. Tatur and L. M.Woods, Strong excitor plasmon coupling in semiconducting carbonnanotube, the entire contents of which are incorporated herein byreference].

Other work has reported about the synthesis and optical properties of acomposite metal-insulator-semiconductor nanowire system which consistsof a wet-chemically grown silver wire core surrounded by a SiO₂ shell ofcontrolled thickness, followed by an outer shell of highly luminescentCdSe nanocrystals [Yuri Fedutik, Vasily Temnov, Ulrike Woggon, ElenaUstinovich, and Mikhail Artemyev Exciton-Plasmon Interaction in aComposite Metal-Insulator-Semiconductor Nanowire System, J. . Am. Chem.Soc., 129 (48), 14939 -14945, 2007, the entire contents of which areincorporated herein by reference]. For a SiO₂ spacer thickness of ˜15nm, they observed an efficient excitation of surface plasmons byexcitonic emission of CdSe nanocrystals. For small d, well below 10 nm,the emission is strongly suppressed (PL quenching), in agreement withthe expected dominance of the dipole-dipole interaction with the dampedmirror dipole [G. W. Ford and W. H. Weber, Electromagnetic interactionsof molecules with metal surfaces,” Phys. Rep. 113, 195-287 (1984), theentire contents of which are incorporated herein by reference]. Fornanowire lengths up to ˜10 μm, the compositemetal-insulator-semiconductor nanowires ((Ag)SiO₂)CdSe act as awaveguide for 1D-surface plasmons at optical frequencies with efficientphoton out coupling at the nanowire tips, which is promising forefficient exciton-plasmon-photon conversion and surface plasmon guidingon a submicron scale in the visible spectral range.

Experiments on colloidal solutions of Ag nanoparticles covered withJ-aggregates demonstrated the possibility of using the strong scatteringcross section and the enhanced field associated with surface plasmon togenerate stimulated emission from J-aggregate excitons with very lowexcitation powers. [Gregory^(,) A. Wurtz, * Paul R. Evans, WilliamHendren, Ronald Atkinson, Wayne Dickson, Robert J. Pollard, and AnatolyV. Zayats, Molecular Plasmonics with Tunable Exciton-Plasmon CouplingStrength in J-Aggregate Hybridized Au Nanorod Assemblies, Nano Lett.,Vol. 7, No. 5, 1297, 2007, the entire contents of which are incorporatedherein by reference]. Their coupling to surface plasmons excitationstherefore provides a particularly attractive approach for creatinglow-powered optical devices. This process can lead to efficient X-raycoupling for phototherapy. In addition, the coupling of J-aggregateswith plasmonics structures presents genuine fundamental interest in thecreation of mixed plasmon-exciton states.

Design, Fabrication and Operation of EPEP Probes

FIG. 21 shows various embodiments of EPEP probes of the inventionshowing the exciton-plasmon coupling:

-   -   (A) probe comprising a PA molecule or group of PA molecules        bound (through a linker, which can be fixed or detachable) to an        energy modulation agent particle that can produce excitons under        radiative excitation at a suitable wavelength (e.g., X-ray). The        energy modulation agent particle is bound to (or in proximity        of) a metal nanoparticle covered with a nanoshell of silica (or        other dielectric material). The silica layer (or nanoshell) (see        FIG. 25A and FIG. 25B; layer nanoshell in white between energy        modulation material and metal nanostructures) is designed to        prevent quenching of the luminescence light emitted by the        energy modulation agent particle excited by X-ray. The metal        nanoparticle (Au, Ag, etc) is designed to induce plasmons that        enhance the X ray excitation that subsequently leads to an        increase in the energy modulation agent light emission,        ultimately enhancing the efficiency of photoactivation, i.e.        phototherapy. The structure of the nanoparticle can also be        designed such that the plasmonics effect also enhances the        energy modulation agent emission light. These processes are due        to strong coupling between excitons (in the energy modulation        agent materials and plasmons in the metal nanoparticles; and    -   (B) probe comprising a PA molecule or group of PA molecules        bound (through a linker, which can be fixed or detachable) to an        energy modulation agent particle that can produce excitons under        radiative excitation at a suitable wavelength (e.g., X-ray). The        energy modulation agent particle is bound to (or in proximity        of) a metal nanoparticle via a spacer (linker). The spacer is        designed to prevent quenching of the luminescence light emitted        by the energy modulation agent particle excited by X-ray.

FIG. 22 shows yet further embodiments of EPEP probes of the invention:

(A) probe comprising a PA molecule or group of PA molecules bound(through a linker, which can be fixed or detachable) to an energymodulation agent particle that can produce excitons under radiativeexcitation at a suitable wavelength (e.g., X-ray). The energy modulationagent particle is covered with a nanoshell of silica (or otherdielectric material), which is covered by a layer of separatenanostructures (nano islands, nanorods, nanocubes, etc . . . ) of metal(Au, Ag). The silica layer (or other dielectric material) is designed toprevent quenching of the luminescence light emitted by the EEC (alsoreferred to as energy modulation agent) particle excited by X-ray. Themetal nanostructures (Au, Ag, etc) are designed to induce plasmons thatenhance the X ray excitation that subsequently leads to an increase inthe EEC light emission, ultimately enhancing the efficiency ofphotoactivation, i.e. phototherapy. The structure of the nanoparticlecan also be designed such that the plasmonics effect also enhance theenergy modulation agent emission light. These processes are due tostrong coupling between excitons (in the energy modulation agentmaterials and plasmons in the metal nanostructures).

(B) probe comprising a group of PA molecules in a particle bound(through a linker, which can be fixed or detachable) to an energymodulation agent particle that can produce excitons under radiativeexcitation at a suitable wavelength (e.g., X-ray). The PA-containingparticle is covered with a layer of metallic nanostructures (Au, Ag).The metal nanostructures (Au, Ag, etc) are designed to induce plasmonsthat enhance the energy modulation agent light emission, ultimatelyenhancing the efficiency of photoactivation, i.e. phototherapy.

(C) probe comprising a PA molecule or group of PA molecules bound(through a linker, which can be fixed or detachable) to an energymodulation agent particle that can produce excitons under radiativeexcitation at a suitable wavelength (e.g., X-ray). The energy modulationagent particle is covered with a nanoshell of silica (or otherdielectric material), which is covered by a layer of metallicnanostructures (Au, Ag). The silica layer (or other dielectric material)is designed to prevent quenching of the luminescence light emitted bythe energy modulation agent particle excited by X-ray. The metalnanostructures (Au, Ag, etc) are designed to induce plasmons thatenhance the X ray excitation that subsequently leads to an increase inthe energy modulation agent light emission, ultimately enhancing theefficiency of photoactivation. In addition. the PA-containing particleis covered with a layer of metallic nanostructures (Au, Ag). The metalnanostructures (Au, Ag, etc) are designed to induce plasmons thatenhance the EEC light emission, ultimately enhancing the efficiency ofphotoactivation.

Hybrid EPEP Nano-Superstructures

EPEP probes can also comprise hybrid self-assembled superstructures madeof biological and abiotic nanoscale components, which can offerversatile molecular constructs with a spectrum of unique electronic,surface properties and photospectral properties for use in phototherapy.

Biopolymers and nanoparticles can be integrated in superstructures,which offer unique functionalities because the physical properties ofinorganic nanomaterials and the chemical flexibility/specificity ofpolymers can be used. Noteworthy are complex systems combining two typesof excitations common in nanomaterials, such as excitons and plasmonsleading to coupled excitations. Molecular constructs comprising buildingblocks including metal, semiconductor nanoparticles (NPs), nanorods(NRs) or nanowires (NWs) can produce EPEP probes with an assortment ofphotonic properties and enhancement interactions that are fundamentallyimportant for the field of phototherapy. Some examples of assemblies ofsome NW nanostructures and NPs have been reported in biosensing.Nanoscale superstructures made from CdTe nanowires (NWs) and metalnanoparticles (NPs) are prepared via bioconjugation reactions.Prototypical biomolecules, such as D-biotin and streptavidin pair, wereutilized to connect NPs and NWs in solution. It was found that Au NPsform a dense shell around a CdTe NW. The superstructure demonstratedunusual optical effects related to the long-distance interaction of thesemiconductor and noble metal nanocolloids. The NW?NP complex showed5-fold enhancement of luminescence intensity and a blue shift of theemission peak as compared to unconjugated NW. [Jaebeom Lee,† AlexanderO. Govorov, John Dulka, and Nicholas A. Kotov, Bioconjugcttes of CdTeNanowires and Au Nanoparticles: Plasmon-Exciton Interactions,Luminescence Enhancement, and Collective Effects, Nano Lett., Vol. 4,No. 12, 2323, 2004, the entire contents of which are incorporated hereinby reference].

FIG. 23 shows various embodiments of EPEP probes of the inventioncomprising superstructures of NPs, NWs and NRs:

(A) probe comprising a PA molecule or group of PA molecules bound(through a linker, which can be fixed or detachable) to an energymodulation agent particle that can produce excitons under radiativeexcitation at a suitable wavelength (e.g., X-ray). The energy modulationagent particle is bound to (or in proximity of) a metal nanowire (ornanorod) covered with a nanoshell cylinder of silica (or otherdielectric material). The silica nanoshells cylinder is designed toprevent quenching of the luminescence light emitted by the energymodulation agent particle excited by X-ray. The metal nanoparticle (Au,Ag, etc) is designed to induce plasmons that enhance the X rayexcitation that subsequently leads to an increase in the energymodulation agent light emission, ultimately enhancing the efficiency ofphotoactivation, i.e. phototherapy. The structure of the nanoparticlecan also be designed such that the plasmonics effect and/or theexciton-plasmon coupling (EPC) effect also enhances the energymodulation agent emission light. These processes are due to strongcoupling between excitons (in the energy modulation agent materials andplasmons in the metal nanoparticles; and

(B) probe comprising a PA molecule or group of PA molecules bound(through a linker, which can be fixed or detachable) to an energymodulation agent particle that can produce excitons under radiativeexcitation at a suitable wavelength (e.g., X-ray). The energy modulationagent particle is bound to (or in proximity of) a metal nanoparticlesvia a spacer (linker). The spacer is designed to prevent quenching ofthe luminescence light emitted by the energy modulation agent particleexcited by X-ray. Same effect as above in (A).

FIGS. 24 and 25 shows another set of embodiments of EPEP probes of theinvention comprising superstructures of NPs, NWs and NRs andbioreceptors (antibodies, DNA, surface cell receptors, etc.). The use ofbioreceptors to target tumor cells has been discussed previously abovein relation to PEPST probes. Note that in this embodiment the PAmolecules are attached along the NW axis in order to be excited by theemitting light form the NWs.

FIG. 26 shows another embodiment of EPEP probes of the inventionincluding superstructures of NPs linked to multiple NWs.

For some embodiments, by adding metal nanostructures designed tointeract specifically with the excitons in the energy modulation agentsystem, there are significant improvements:

(1) an additional radiative pathway from exciton to photon conversion isintroduced

(2) the metal nanostructures can be designed to amplify (due to theplasmonics effect) the excitation radiation (e.g., X-ray) and/or theemission radiation (e.g, UV or visible) to excite the photo-active (PA)molecule, thereby enhancing the PA effectiveness.

Various metallic nanostructures that can be used in EPEP probeembodiments of the invention are the same as those illustrated in FIG. 4for the PEPSI probes.

EPEP Probes with Microresonators

In one embodiment, the energy modulation agent system can be designed toserve also as a microresonator having micron or submicron size. Priorwork has described a resonant microcavity and, more particularly, to aresonant microcavity which produces a strong light-matter interaction [WLipson; L.C. Kinierling; Lionel C, Resonant microcavities, U.S. Pat. No.6,627,923, 2000, the entire contents of which are incorporated herein byreference]. A resonant microcavity, typically, is formed in a substrate,such as silicon, and has dimensions that are on the order of microns orfractions of microns. The resonant microcavity contains optically-activematter (i.e., luminescent material) and reflectors which confine lightin the optically-active matter. The confined light interacts with theoptically-active matter to produce a light-matter interaction. Thelight-matter interaction in a microcavity can be characterized as strongor weak. Weak interactions do not alter energy levels in the matter,whereas strong interactions alter energy levels in the matter. In stronglight-matter interaction arrangements, the confined light can be made toresonate with these energy level transitions to change properties of themicrocavity.

Experimental Methods

Preparation of Nanoparticles (Ag, Au)

There are numerous methods to prepare metal nanoparticles for EPEP orPEPST probes. Procedures for preparing gold and silver colloids includeelectroexplos on, electrodeposition, gas phase condensation,electrochemical methods, and solution-phase chemical methods. Althoughthe methodologies for preparing homogeneous-sized spherical colloidalgold populations 2-40 nm in diameter are well known [N. R. Jana, L.Gearheart and C. J. Murphy, Seeding growth for size control of 5-40 nmdiameter gold nanoparticles. Langmuir 17 (2001), pp. 6782-6786, theentire contents of which are incorporated herein by reference], andparticles of this size are commercially available. An effective chemicalreduction method for preparing populations of silver particles (withhomogeneous optical scattering properties) or gold particles (withimproved control of size and shape monodispersity) is based on the useof small-diameter uniform-sized gold particles as nucleation centers forthe further growth of silver or gold layers.

A widely used approach involves citrate reduction of a gold salt toproduce 12-20 nm size gold particles with a relatively narrow sizedistribution. One commonly used method for producing smaller goldparticles is described in Brust, M; Walker, M.; Bethell, D.; Schiffrin,D. I; Whyman, R. Chem. Commun. 1999, 801, the entire contents of whichare incorporated herein by reference. This method is based onborohydride reduction of gold salt in the presence of an alkanethiolcapping agent to produce 1-3 nm particles. Nanoparticle sizes can becontrolled between 2 and 5 nm by varying the thiol concentration,[Hostetler, M. J.; Wingate, J. E.; Zhong, C. J.; Harris, J. E.; krachet,R. W.; Clark, M. R.; Londono, J. D.; Green, S. J.; Stokes, J. J.;Wignall, G. D.; Glish, G. L.; Porter, M D.; Evans, N. D.; Murray, R. W.Langmuir 1998. 14, 17, the entire contents of which are incorporatedherein by reference]. Phosphine-stabilized gold clusters have also beenproduced and subsequently converted to thiol-capped clusters by ligandexchange in order to improve their stability [Schmid, G.; Pfeil, R.;Boese, R.; Bandrmann, F.; Meyer, S.; Calls, G. H. M; van der Velden, J.W. A. Chem. Ber. 1981, 114, 3634; Warner, M. G.; Reed, S. M; Hutchison,J. E. Chem. Mater. 2000,12, 3316, the entire contents of which areincorporated herein by reference] and phosphine- stabilizedmonodispersed gold particles were prepared using a similar protocol tothe Brust method [Weare, W. W.; Heed, S. M.; Warner, M. G.; Hutchison,J. E. J. Am. Chem. Soc. 2000, 122, 12890, the entire contents of whichare incorporated herein by reference]. See also: Ziyi Zhong, BenoitMale, Keith B. Luong, John H. T., More Recent Progress in thePreparation of Au Nanostructures, Properties, and Applications,Analytical Letters; 2003, Vol. 36 Issue 15, p3097-3118, the entirecontents of which are incorporated herein by reference.

Fabrication of Nanoparticle of Metal Coated with Nanoshells of Dyes Thefabrication of metal nanoparticles coated with nanoshells of dyemolecules can be performed using the method described in Akito Masuhara,Satoshi Ohhashi, Hitoshi Kasai; Shuji Okada, FABRICATION AND OPTICALPROPERTIES OF NANOCOMPLEXES COMPOSED OF METAL NANOPARTICLES AND ORGANICDYES, Journal of Nonlinear Optical Physics & Materials Vol. 13, Nos. 3 &4 (2004) 587-592, the entire contents of which are incorporated hereinby reference. Nanocomplexes composed of Ag or Au as a core and3-carboxlymethyl-5-[2-(3- octadecyl-2-benzoselenazolinylidene)ethylidene]rhodanine (MCSe) or copper (II) phthalocyanine (CuPc) as ashell are prepared by the co-reprecipitation method. In the case ofAg-MCSe nanocomplexes, 0.5 mM acetone solution of MCSe are injected into10 ml of Ag nanoparticle water dispersion, prepared by the reduction ofAgNO₃ using NaBH₄: Au-MC Se nanocomplexes are also fabricated in asimilar manner. A water dispersion of Au nanoparticles was prepared bythe reduction of HAuCl₄ using sodium citrate. Subsequently, 2 M NH₄OH(50 ul) was added and the mixture was thermally treated at 50° C. Thisamine treatment often stimulates the J-aggregate formation of MCSe.6Ag-CuPc and Au-CuPc nanocomplexes were also fabricated in the samemanner: 1 mM 1-methyl-2-pyrrolidinone (NMP) solution of CuPc (200 μl)was injected into a water dispersion (10 ml) of Ag or Au nanoparticles.

Preparation of silver nanoparticles

Silver (or gold) colloids were prepared according to the standardLee-Meisel method: 200 mL of 10⁻³ M AgNO₃ aqueous solution was boiledunder vigorous stirring, then 5 mL of 35-mM sodium citrate solution wereadded and the resulting mixture was kept boiling for 1 h.

This procedure was reported to yield ˜10¹¹ particles/mL of homogenouslysized colloidal particles with a diameter of ˜35-50 nm and an absorptionmaximum at 390 nm. The colloidal solutions were stored at 4° C. andprotected from room light. Further dilutions of the colloidal solutionswere carried out using distilled water.

Fabrication/Preparation of Metal Nanocaps

One approach has involved the use of nanospheres spin-coated on a solidsupport in order to produce and control the desired roughness. Thenanostructured support is subsequently covered with a layer of silverthat provides the conduction electrons required for the surface plasmonmechanisms. Among the techniques based on solid substrates, the methodsusing simple nanomaterials, such as Teflon or latex nanospheres, appearto be the simplest to prepare. Teflon and latex nanospheres arecommercially available in a wide variety of sizes. The shapes of thesematerials are very regular and their size can be selected for optimalenhancement. These materials comprise isolated dielectric nanospheres(30-nm diameter) coated with silver producing systems ofhalf-nanoshells, referred to as nanocaps.

Fabrication of Gold Nanoshells

Gold nanoshells have been prepared using the method described in HirschL R, Stafford R J, Bankson J A, Sershen S R, Price R E, Hazle J D, HalasN J, West J L (2003) Nanoshell-mediated near infrared thermal therapy oftumors udder MR Guidance. Proc Natl Acad Sci 100:13549-13554. Thismethod uses a mechanism involving nucleation and then successive growthof gold nanoparticles around a silica dielectric core. Goldnanoparticles, the seed, prepared as described above using the Frensmethod, were used to grow the gold shell. Silica nanoparticles (100nm)used for the core of the nanoshells were monodispersed in solution of 1%APTES in EtOH. The gold “seed” colloid synthesized using the Frensmethod were grown onto the surface of silica nanoparticles via molecularlinkage of amine groups. The “seed” covers the aminated silicananoparticle surface, first as a discontinuous gold metal layergradually growing folnling a continuous gold shell.

Commercial Applications In the following commercial applications of theinvention described here, the energy modulation agents 3 (e.g.,luminescing particles or photon emitters) are provided and distributedinto a medium 4 for deactivation or activation of agents in the mediumto produce a physical, chemical, or biological change in the medium. Inone embodiment, plasmonics agents as described above are added to themedium. The plasmonics agents can enhance both the applied initiationenergy such that the enhanced initiation energy activates the at leastone activatable agent which produces a change in the medium whenactivated and can enhance light converted by the energy modulationagents.

Examples of luminescing particles can include gold particles (such asfor example the nanoparticles of gold described above), BaFBr:Euparticles, CdSe particles, Y₂O₃:Eu³⁺ particles, and/or other knownstimulated luminescent materials such as for example ZnS: Mn²⁺; ZnS:Mn²⁺,Yb³⁺, Y₂ O₃: EU³⁺; BaFBr:Tb³⁺; and YF₃:Tb³+.

In one embodiment of the invention described here, other potentiallyuseful luminescing particles (or energy modulation agents) includecarbon nanotubes as described for example by Wang et al. in“Electromagnetic excitation of nano-carbon in vacuum,” OPTICS EXPRESS,Vol 13, No. 10, May 10, 2005, the entire contents of which areincorporated herein by reference. Such carbon nanotubes show both blackbody emission and discrete line-type emissions in the visible whenexposed to microwave irradiation.

Other potentially useful luminescing particles for the inventiondescribed here include the chemiluminescent reactions/species describedby Asian et al. in “Multicolor Microwave-Triggered Metal-EnhancedChemiluminescence,” in J. AM. CHEM. SOC. published on Web Sep. 23, 2006,the entire contents of which are incorporated herein by reference. Thesechemiluminescent reactions/species are formed with silver nanoparticleswhich enhance the chemiluminescent reactions when exposed to microwaveradiation. Aslan et al. utilized chemiluminescent species fromcommercial glow sticks where for example hydrogen peroxide oxidizesphenyl oxalate ester to a peroxyacid ester and phenol. The unstableperoxyacid ester decomposes to a peroxy compound and phenol, the processchemically inducing an electronic excited state responsible for thelight emission. While these chemiluminescent species will have a limitedlifetime, there use in curing applications for the invention describedhere is still viable where the cure process is a one-time occurrence,and the external microwave source accelerates the cure by acceleratedvisible light production.

The luminescent wavelength and/or efficiency of the luminescentparticles often depend on the size of the particle. Particle sizes inthe nanometer size range for the invention described here exhibitstronger luminescence in many cases, as described in U.S. Pat. Appl.Publ. No. 2007/0063154, whose entire contents are incorporated herein byreference. Further, in one embodiment of the invention described here,the luminescing particles can be combined with molecular complexes suchas poly(ethylene glycol), vitamin B12, or DNA, which serves to mitigateagainst coagulation of the luminescing particles (especially thenanoparticles) and serves to make the luminescing particlesbiocompatible. More specifically, one recipe for the synthesis of CdSenanocrystals is given here from U.S. Pat. Appl. Publ. No. 2007/0063154.Accordingly, citrate-stabilized CdSe nanocrystals suitable for theinvention described here can be prepared according to the followingprocedure:

-   -   To 45 ml of water are added 0.05 g sodium citrate (Fluka) and 2        ml of 4×10 ⁻² M cadmium perchlorate (Aldrich). The pH is        adjusted to 9.0 by 0.1 M NaOH (Alfa). The solution is bubbled        with nitrogen for 10 minutes, and then 2 ml of lx 10 ⁻² M        N,N-dimethylselenourea (Alfa) is added. The mixture is heated in        a conventional 900-watt microwave oven for 50 seconds. In this        recipe, the Cd:Se molar ratio is 4:1, which leads to CdSe        nanoparticles with ^(˜)4.0 nm diameter; by increasing the Cd        concentration it is possible to synthesize smaller CdSe        nanoparticles.

Further, the luminescing particles for the invention described here canbe coated with insulator materials such as for example silica which willreduce the likelihood of any chemical interaction between theluminescing particles and the medium. For biological applications ofinorganic nanoparticles, one of the major limiting factors is theirtoxicity. Generally speaking, all semiconductor nanoparticles are moreor less toxic. For biomedical applications, nanoparticles with toxicityas low as possible are desirable or else the nanoparticles have toremain separated from the medium. Pure TiO₂ , ZnO, and Fe₂O₃ arebiocompatible. CdTe and CdSe are toxic, while ZnS, CaS, BaS, SrS and Y₂O₃ are less toxic. In addition, the toxicity of nanoparticles can resultfrom their inorganic stabilizers, such as TGA, or from dopants such asEu²⁺, Cr³⁺ or Nd ³⁺. Other suitable energy modulation agents which wouldseem the most biocompatible are zinc sulfide, ZnS.Mn²⁺, ferric oxide,titanium oxide, zinc oxide, zinc oxide containing small amounts of Al₂O₃and AgI nanoclusters encapsulated in zeolite. For non-medicalapplications, where toxicity may not be as critical a concern, thefollowing materials (as well as those listed elsewhere) are consideredsuitable: lanthanum and gadolinium oxyhalides activated with thulium;Er³⁺ doped BaTiO₃ nanoparticles, Yb³⁺ doped CsMnCl₃ and RbMnCl₃,BaFBr:Eu²⁺ nanoparticles, Cesium Iodine, Bismuth Germanate, CadmiumTungstate, and CsBr doped with divalent Eu.

In various embodiments of the invention, the following luminescentpolymers are also suitable as energy modulation agents: poly(phenyleneethynylene), poly(phenylene vinylene), poly(p-phenylene),poly(thiophene), poly(pyridyl vinylene), poly(pyrrole), poly(acetylene),poly(vinyl carbazole), poly(fluorenes), and the like, as well ascopolymers and/or derivatives thereof.

While many of the energy modulation agents of the invention are downconversion agents (i.e. where higher energy excitation produces lowerenergy emission), U.S. Pat. No. 7,008,559 (the entire contents of whichare incorporated herein by reference) describes the upconversionperformance of ZnS where excitation at 767 nm produces emission in thevisible range. The materials described in U.S. Pat. No. 7,008,559including the ZnS as well as Er³⁺doped BaTiO₃ nanoparticles and Yb³⁺doped CsMnCl₃ are suitable in various embodiments of the invention.

Further materials specified for up conversion include CdTe, CdSe, ZnO,CdS, Y₂O₃, MgS, CaS, SrS and BaS. Such up conversion materials may beany semiconductor and more specifically, but not by way of limitation,sulfide, telluride, selenide, and oxide semiconductors and theirnanoparticles, such as Zn_(1-x)Mn_(x)S_(y), Zn_(1-x)Mn_(x)Se_(y),Zn_(1−x)Te_(y), Cd_(1-x)MnS_(y), Cd_(1-x)Mn_(x)Se_(y),Cd_(1-x)Mn_(x)Te_(y), Pb_(1-x)Mn_(x)S_(y), Pb_(1-x)Mn_(x)Se_(y),Pb_(1-x)Mn_(x)Te_(y), Ca_(1-x)Mn_(x)S_(y), Ba_(1-x)Mn_(x)S_(y) andSr_(1-x), etc. (wherein, 0<x≤1, and 0<y≤1). Complex compounds of theabove-described semiconductors are also contemplated for use in theinvention—e.g. (M_(1-z)N_(z))_(1-x)Mn_(x)A_(1−y)B_(y)Mn═Zn, Cd, Pb, Ca,Ba, Sr, Mg; N=Zn, Cd, Pb, Ca, Ba, Sr, Mg; A═S, Se, Te, O; B=S, Se, Te,O; 0<x≤1, 0<y≤1, 0<z≤1). Two examples of such complex compounds areZn_(0.4)Cd_(0.4)Mn_(0.2)S and Zn_(0.9)Mn_(0..1)S_(0.8)Se_(0.2).Additional conversion materials include insulating and nonconductingmaterials such as BaF₂, BaFBr, and BaTiO₃, to name but a few exemplarycompounds. Transition and rare earth ion co-doped semiconductorssuitable for the invention include sulfide, telluride, selenide andoxide semiconductors and their nanoparticles, such as ZnS; Mn; Er; ZnSe;Mn, Er; MgS; Mn, Er; CaS; Mn, Er; ZnS; Mn, Yb; ZnSe; Mn,Yb; MgS; Mn, Yb;CaS; Mn,Yb etc., and their complex compounds:(M_(1-z)N_(z))_(1-x)(MnR_(1-q))_(x)A_(1-y)B_(y) (M═Zn, Cd, Pb, Ca, Ba,Sr, Mg; N═Zn, Cd, Pb, Ca, Ba, Sr, Mg; A═S, Se, Te, O; B═S, . . . 0<z≤1,o<q≤1).

Indeed, some nanoparticles such as ZnS:Tb³⁺, Er³⁺; ZnS:Tb³⁺; Y₂O₃:Tb³⁺;Y₂O3:Tb³⁺, Er³⁺; ZnS:Mn²⁺; ZnS:Mn,Er³⁺are known in the art to have twofunctions, capable of functioning for both down-conversion luminescenceand upconversion luminescence.

To reduce the toxicity or to make these nanoparticles bio-inert orbiocompatible, one embodiment of the invention described here coatsthese nanoparticles with silica. Silica is used as a coating material ina wide range of industrial colloid products from paints and magneticfluids to high-quality paper coatings. Further, silica is bothchemically and biologically inert and also is optically transparent. Inthe following recipe (from Al. A. Correa-Duarte, M. Giesig, and L. M.Liz-Marzan, Stabilization of CdS semiconductor nanoparticles againstphotoclegradation by a silica coating procedure, Chem. Phys. Lett.,1998, 286: 497, the entire contents of which is explicitly incorporatedherein by reference in its entirety), citrate-stabilized CdTe:Mn 2+/SiO₂nanocrystals suitable for the invention described here can be preparedwith a silica coating:

-   -   (1) To a CdTe:Mn 2+ nanoparticle solution (50 ml), a freshly        prepared aqueous solution of 3-(mercaptopropyl) trimethoxysilane        (MPS) (0.5 ml, 1 mM) (Sigma) is added under vigorous stirring.        The function of MPS is that its mercapto group can directly bond        to the surface Cd sites of CdTe, while leaving the silane groups        pointing toward solution from where silicate ions approach the        particle surface; (2) Addition of 2 ml of sodium silicate (Alfa)        solution at pH of 10.5 under vigorous stirring; (3) The        resulting dispersion (pH ^(˜)8.5) is allowed to stand for 5        days, so that silica slowly polymerizes onto the particle        surface; and (4) Transfer of the dispersion to ethanol so that        the excess dissolved silicate can precipitate out, increasing        the silica shell thickness.

Alternatively, as shown in FIG. 3C and FIG. 3D, luminescing particles inencapsulated structures 10 could be placed in the vicinity of themedium. In one embodiment for the invention described here, luminescingparticles are coated on the interior of quartz or glass tubes 9 andsealed. In another embodiment, luminescing particles could be coated onthe surface of spheres or tubes, and afterwards encapsulated with silica(or other suitable passivation layer) using a vapor deposition orsputtering process or spin-on glass process of the solution processdescribed above to make the encapsulation structures 10 which may bepart of re-entrant structures extending from walls of a container (as inFIG. 3C) or which may be part of a fluidized bed structure (as in FIG.3D). In another embodiment, the plasmonics agents are fixed to an outersurface of the glass tubes 9. External light applied to the tubes andscattered to the outer surfaces is enhanced at the plasmonics agentspermitting more efficient treatment of the medium without necessarilyhaving to use energy modulation agents.

In the either configuration, the medium to be treated would flow by theencapsulated structures 10, or flow along with encapsulated structures6, and the separation distance between the encapsulated structures 6, 10would be set a distance smaller than the UV penetration depth in themedium.

A suitable light source (such as one of the x-ray sources discussedabove) can be used to stimulate the luminescing particles in theencapsulated structures 10. In one embodiment of the invention describedhere, the concentration of luminescing particles in the medium or thespacing between the encapsulated structures 10 is set such thatluminescing particles are separated from each other in the medium byless than a UV depth of penetration into the medium. Higherconcentrations are certainly usable and will generate higher UV fluxesshould the energy source have enough intensity to “light” all theluminescing particles.

For a relatively unclouded aqueous medium, solar UV-B irradiancedecreases to 1% after penetration into the water samples between 0.2 mand 1 m, whereas UV-A penetrates on the order of several meters. Forsuch mediums, the concentration of luminescing particles is moredetermined by the time needed for the intended UV flux to producedeactivation or activation of an agent in the medium, rather than havingto be set based on a concentration of luminescent particles where themedium itself does not occlude the UV stimulated emission frompenetrating throughout the medium. The placement of the luminescentparticles in the medium and in the vicinity of the medium is notrestricted by the optical density of the medium.

Based on published data of an average of 5.2 spontaneous photons emittedfrom BaFBr:Eu ²⁺ for every keV of X-ray absorbed (M. Thorns, H. vonSeggern, Method for the determination of photostimulable defect centerconcentrations, production rates, and effective formation energies, J.Appl. Phys. 1994, 75: 4658-4661, the entire contents of which is hereinexplicitly incorporated by reference in its entirety.), one expects thatabout 50 photons are emitted from a CdTe nanoparticle for each 50 keVX-ray absorbed.

Based on the results in U.S. Pat. Appl. Publ. No. 2007/0063154 for X-rayspectra of CdTe/BaFBr:Eu ²⁺ nanocomposites prepared using aconcentration of 0.8 ml L-cysteine stabilized CdTe particle solution in0.2 g BaFBr:Eu ²⁺phosphor. As the X-ray irradiation time increases, theX-ray luminescence intensity of Eu ²⁺at 390 nm increases in intensity.This phenomenon has been discussed in W. Chen, S. P. Wang, S. Westcott,J. Zhang, A. G. Joly, and D. E. McCready, Structure and luminescence ofBaFBr:Eu²⁺ and BaFBr:Eu ²⁺, Tb³⁺ phosphors and thin films, J. Appl.Phys. 2005, 97: 083506, the entire contents of these references areherein incorporated by reference in their entirety.

Hence, in one embodiment of the invention, a minimum baselineconcentration of about 10⁹ nanoparticles per cm³ for 200 nm diameterparticles is expected to be sufficient for UV emission to produce achange in the medium. The invention is not limited to this concentrationrange, but rather this range is given as an illustrative example.Indeed, higher concentrations will increase the UV emission per unittime and provide faster reactions, which in general would be consideredmore useful in industrial applications where product throughput is aconcern.

Sterilization and Cold Pasteurization of Fluids

Table 1 included below shows appropriate intensities for germicidaldestruction.

TABLE 1 Germicidal energies needed to destroy Approximate intensity(μW/cm²) required for 99% destruction of microorganisms: Bacteria 10 400Protozoa (single celled organism) 105 000  Paramecium (slipper shaped200 000  protozoa) Chlorella (unicellular fresh-water 13 000 alga)Flagellate(protozoan or alga with 22 000 flagella) Sporozoan (parasiticprotozoans) 100 000  Virus  8 000

Accordingly, the energy modulation agents (or luminescing particles) ofthe invention as discussed above with regard to FIGS. 3B and 3C) can beprovided on the interior of sealed quartz or glass tubes or can beprovided coated on the surface of spheres or tubes, and furtherencapsulated with a silica or passivation layer. Plasmonics agents canbe formed with the energy modulation agents. In either configuration forthe invention described here, a medium could flow by the encapsulatedstructures 6, 10 with a separation distance between the encapsulatedstructures or the quartz or glass tubes being made smaller than the UVpenetration depth.

For example, it is known that ultraviolet (UV) with a wavelength of 254nm tends to inactivate most types of microorganisms. Most juices areopaque to UV due to the high-suspended solids in them and hence theconventional UV treatment, usually used for water treatment, cannot beused for treating juices. In order to make the process efficient, a thinfilm reactor constructed from glass has been used with the juice flowingalong the inner surface of a vertical glass tube as a thin film. See“Ultraviolet Treatment of Orange Juice” by Tran et al. published inInnovative Food Science & Emerging Technologies (Volume 5, Issue 4,December 2004, Pages 495-502), the entire contents of which areincorporated herein by reference. Tran et al. reported therein decimalreduction doses required for the reconstitute orange juices (OJ; 10.5°Brix) were 87+7 and 119+17 mJ/cm² for the standard aerobic plate count(APC) and yeast and moulds, respectively. In that article, the shelflife of fresh squeezed orange juice was extended to 5 days with alimited exposure of UV (73.8 mJ/cm²). The effect of UV on theconcentration of Vitamin C was investigated using both HPLC andtitration methods of measurements. The degradation of Vitamin C was 17%under high UV exposure of 100 mJ/cm², which was similar to that usuallyfound in thermal sterilization. Enzyme pectin methylesterase (PME)activity, which is the major cause of cloud loss of juices, was alsomeasured. The energy required for UV treatment of orange juice (2.0 kWh/m³) was much smaller than that required in thermal treatment (82 kWh/m³). The color and pH of the juice were not significantly influencedby the treatment.

The invention described herein offers advantages over this approach inthat the energy modulation agents can be placed inside fixtures such asquartz or glass (encapsulation structures 8) within the orange juice (orother fluid medium) and irradiated with x-rays (or other penetratingradiation) through for example a plastic or aluminum container 9 toactivate the energy modulation agents 3 and 6 in the orange juice. Assuch, the expense and fragility of a thin film reactor constructed fromglass of other similar structure is avoided.

While discussed with regard to orange juice, any other medium to besterilized including food products, medical products and cosmeticproducts could be treated using the technique of the invention describedherein.

Sterilization of Medical and Pharmaceutical Articles

As noted above, medical bottle caps need to be sterilized between thebase cap material and the seal material which contacts to the base ofthe medical bottle. Steam autoclaves are insufficient for this purposeas once glued, the steam is unable to penetrate into the glue seam.

Gamma irradiation has been used conventionally to sterilize medicalbottle caps and other medical, pharmaceutical, and cosmetic articlessuch as surgical disposables (e.g., surgical bandages, dressings, gaugepads, nappies, delivery kits, and etc.), metallic products (e.g.,surgical blades, implants, aluminum caps, containers, etc.), and plasticand rubber Items(e.g., petri-dish, centrifuge tube, blood collectionsets, scalp vein sets, shunt valves, rubber gloves, contraceptivedevices, gowns, wraps covers, sheets, etc.).The invention would beapplicable for the sterilization of any “interior” surfaces of these andother products.

In one embodiment of the invention described herein, UV luminescentparticles would be included in an adhesive layer when the seal materialis applied to the bottle cap. X-ray irradiation would then be capable ofcuring the adhesive (if for example the adhesive were a photosensitiveadhesive as discussed below in greater detail) and would produce withinthe adhesive medium UV radiation for direct sterilization or for theproduction of singlet oxygen or ozone for biological germicide.Additionally, plasmonics agents can be included to enhance the effect ofthe incident radiation or the internally generated radiation.

While illustrated here with regard to medical bottle caps, otheradhesively constructed devices could benefit from these procedures inwhich the adhesive medium is cured and/or sterilized during activationof energy modulation agents 3 and 6.

Sterilization of Blood Products

U.S. Pat. No. 6,087,141 (the entire contents of which are incorporatedherein by reference) describes an ultraviolet light actived psoralenprocess for sterilization of blood transfusion products. Here, theinvention can be applied for example in the equipment shown in FIGS. 3Cand 3D for the treatment of orthe neutralization of AIDS and HIV orother viral or pathogenic agents in blood transfusion products. In thisembodiment, at least one photoactivatable agent is selected frompsoralens, pyrene cholesteryloleate, acridine, porphyrin, fluorescein,rhodamine, 16-diazorcortisone, ethidium, transition metal complexes ofbleomycin, transition metal complexes of deglycobleomycin organoplatinumcomplexes, alloxazines, vitamin Ks, vitamin L, vitamin metabolites,vitamin precursors, naphthoquinones, naphthalenes, naphthols andderivatives thereof having planar molecular conformations,porphorinporphyrins, dyes and phenothiazine derivatives, coumarins,quinolones, quinones, and anthroquinones. These photoactivatable agentsare introduced into the blood product(or a patient's blood stream). Apenetrating energy is applied to the blood product (or to the patient).The energy modulation agents (either included in the blood product) orin encapsulated structures 10 generate secondary light such as UV lightwhich activates the photoactivatable agents in the blood products.

In a specific example, the photoactivatable agent is a psoralen, acoumarin, or a derivative thereof, and as discussed above, one cansterilize blood products in vivo (i.e., in a patient) or in a containerof the blood product (such as for example donated blood). The treatmentcan be applied to treat disorders such as for example a cancer cell, atumor cell, an autoimmune deficiency symptom virus, or a blood-bornegermicide is treated by the psoralen, the coumarin, or the derivativethereof.

Waste Water Detoxification

Photocatalysis has also been used as tertiary treatment for wastewaterto comply with the regulatory discharge limits and to oxidize persistentcompounds that have not been oxidized in the biological treatment.Photocatalysis has being applied to the elimination of severalpollutants (e.g., alkanes, alkenes, phenols, aromatics, pesticides) withgreat success. In many cases, total mineralization of the organiccompounds has been observed. Several photocatalysts, such as CdS, Fe₂O₃,ZnO, WO₃, and ZnS, have been studied, but the best results have beenachieved with TiO₂ P₂₅. These photocatalyst are usable for the inventiondescribed here.

The wastewaters of an oil refinery are the waters resulting from washingthe equipment used in the process, undesirable wastes, and sanitarysewage. These effluents have high oil and grease contents, besides otherorganic compounds in solution. These pollutants form a residual chemicaloxygen demand (COD) that may pose serious toxic hazards to theenvironment.

It is known that photocatalysis can be used for waste water reductionremediation. U.S. Pat. No. 5,118,422 (the entire contents of which areincorporated herein by reference) to Cooper et al. describe anultraviolet driven photocatalytic post-treatment technique for purifyinga water feedstock containing an oxidizable contaminant compound. In thiswork, the water feedstock was mixed with photocatalytic semiconductorparticles (e.g., TiO₂, ZnO, CdS, CdSe, SnO₂, SrTiO₃, WO₃, Fe₂O₃, andTa₂O₅ particles) having a particle size in the range of about 0.01 toabout 1.0 micron and in an amount of between about 0.01% and about 0.2%by weight of the water. The water including the semiconductor mixture isexposed to band-gap photons for a time sufficient to effect an oxidationof the oxidizable contaminant to purify the water. Crossflow membranefiltration was used to separate the purified water from thesemiconductor particles. Cooper et al. show that the organic impuritycarbon content of simulated reclamation waters at nominal 40 PPM levelwere reduced to parts per billion using a recirculation batch reactor.

Cooper el al. identified that one important aspect of the photocatalyticprocess is the adsorption of the organic molecules onto the extremelylarge surface area presented by the finely divided powders dispersed inthe water. Cooper et al. further indicated that, in photoelectrochemicalapplications, advantage is taken of the fact that the solid phase (ametal oxide semiconductor) is also photo-active and that the generatedcharge carriers are directly involved in the organic oxidation. Theadsorption of the band-gap photon by the semiconductor particle resultsin the formation of an electron (e⁻)/hole(h⁺) pair. Cooper et al.explain that the electrons generated in the conduction band react withsolution oxygen forming the dioxygen anion (O²⁻) species whichsubsequently undergo further reactions resulting in the production ofthe powerfully oxidizing hydroxyl radical species, OH. These powerfuloxidants are known to oxidize organic compounds by themselves.Additionally, Cooper et al. explain that the strongly oxidizing holesgenerated in the valence band have sufficient energy to oxidize allorganic bonds.

In the reactor of Cooper et al., turbulence is necessary in order toensure that the waste water contaminants and the photocatalytic titan aparticles are exposed to the UV light. Cooper et al. explain that themost basic considerations of photocatalyst light adsorption and itsrelationship to convective mixing. For a 0.1 wt % photocatalyst loading,experiments have shown that 90% of the light is absorbed within 0.08 cm.This is primarily due to the large UV absorption coefficient of thephotocatalyst and therefore, most of the photoelectrochemistry occurswithin this illuminated region. By operating the reactor of Cooper etal. with a Reynolds number (Re) of 4000, a significant portion of thephotoactive region is ensured of being within the well mixed turbulentzone.

Santos et al. have reported in “Photocatalysis as a tertiary treatmentfor petroleum refinery wastewaters” published in Braz. J. Chem. Eng.vol. 23, No. 4, 2006 (the entire contents of which are incorporatedherein by reference), photocatalysis for tertiary treatment forpetroleum refinery wastewaters which satisfactorily reduced the amountof pollutants to the level of the regulatory discharge limits andoxidized persistent compounds that had not been oxidized in thebiological treatment. The treatment sequence used by the refinery(REDUC/PE1ROBRAS, a Brazilian oil refinery) is oil/water separationfollowed by a biological treatment. Although the process efficiency interms of biological oxygen demand (BOD) removal is high, a residual andpersistent COD and a phenol content remains. The refining capacity ofthe refinery is 41,000 m³/day, generating 1,100 m³/h of wastewater,which are discharged directly into the Guanabara Bay (Rio de Janeiro).Treating the residual and persistent COD remains a priority.

Santos et al. conducted a first set of experiments carried out in anopen 250 mL reactor containing 60 mL of wastewater. In the second set ofexperiments, a Pyrex® annular reactor containing 550 mL of wastewaterwas used (De Paoli and Rodrigues, 1978), as shown in FIG. 1. Thereaction mixtures inside the reactors were maintained in suspension bymagnetic stirring. In all experiments, air was continuously bubbledthrough the suspensions. A 250 W Phillips HPL-N medium pressure mercuryvapor lamp (with its outer bulb removed) was used as the UV-light source(radiant flux of 108 J·m⁻²·s⁻¹ at 8>254 nm). In one set of experiments,the lamp was positioned above the surface of the liquid at a fixedheight (12 cm). In the second set, the lamp was inserted into the well.All experiments by Santos et al. were perfoi tried at 25±1° C. Thecatalyst concentration ranged from 0.5 to 5.5 g L⁻¹ and the initial pHranged from 3.5 to 9.

In the invention described herein, luminescing particles or other energymodulation agents would be placed inside quartz or glass fixtures withinthe waste water or would be placed on silica encapsulated structureswithin the waste water which, like the photocatalytic TiO₂, could beentrained in the waste water during the irradiation. Additionally, theplasmonics agents can be included to enhance the effect of the incidentradiation or the internally generated radiation.

Upon irradiation with x-rays (or other penetrating radiation) throughfor example a plastic or aluminum container, activation of theluminescing particles (i.e., energy modulation agents) would generate UVlight in nearby presence of the photocatalytic agent. In other words forthe invention described herein, the luminescent particles or otherenergy modulation agents are mixed along with the photocatalyticsemiconductor particles in the waste water fluid stream, and theexterior activation energy source penetrates the container (e.g., aplastic or aluminum container) and irradiates the bulk of the wastewater, producing UV light throughout the waste water which in turndrives the photocatalytic reactions. In one embodiment, the plasmonicsagents are complexed with the luminescent particles or other energymodulation agents prior to being added to the fluid stream.

As such, the invention described herein offers a number to advantagesover that described above, including the elimination of expensiveholding tanks for the waste water, the avoidance of having to pump thewastewater at higher pressures or flowrates to produce sufficientturbulence, and the generation of UV light throughout the wastewater tothereby provide faster bulk processing of the waste water.

Photostimulation

Photostimulation is a field in which light is applied to in order toalter or change a physical property. For example, there has been anincreased focus on the use of biodegradable polymers in consumer andbiomedical fields. Polylactic acid (PLA) plastics andpolyhydroxyalkanoates (PHA) plastics have been playing a vital role infulfilling the objectives. But their relatively hydrophobic surfaceslimit their use in various applications. Hence, there is a need tosurface modify these film surfaces. Due to the lack of any modifiableside chain groups, workers have used a sequential two step photograftingtechnique for the surface modification of these biopolymers. In stepone, benzophenone was photografted on the film surface and in step two,hydrophilic monomers like acrylic acid and acrylamide werephotopolymerized from the film surfaces.

Workers have found that UV irradiation could realize an effective graftcopolymerization. UV-assisted photografting in ethanol has been used togrow hydrophilic polymers (e.g., poly(acrylic acid) and polyacrylamide)from the surfaces of PLA, PHA, and PLA/PHA blend films. In that work, afunctional polyurethane (PU) surface was prepared by photo-graftingN,N-dimethylaminoethyl methacrylate (DMAEM) onto the membrane surface.Grafting copolymerization was conducted by the combined use of thephoto-oxidation and irradiation grafting. PU membrane was photo-oxidizedto introduce the hydroperoxide groups onto the surface, then themembrane previously immersed in monomer solution was irradiated by UVlight. Results have shown prior to the invention that UV irradiation canrealize graft copolymerization effectively.

In the invention described herein, these processes are expedited by theinclusion of luminescing particles or other energy modulation agents indispersion in the fluid medium being used for photostimulation.Additionally, the plasmonics agents can be included to enhance theeffect of the incident radiation or the internally generated radiation.In one embodiment, the plasmonics agents are complexed with theluminescent particles or other energy modulation agents prior to beingadded to the fluid medium.

Upon irradiation with x-rays (or other penetrating radiation) throughfor example a plastic or aluminum container, activation of theluminescing particles (i.e., energy modulation agents) would generate UVlight throughout the volume of the medium (eliminating any shadowingeffects) and permitting batch or bulk type processing to occur inparallel throughout the container.

In other examples, the interior generation of light inside a bulk mediummay serve to stimulate a chemical or biological process either by directinteraction of the light with activatable agents in the medium or theindirect generation of heat which the invention described here by way ofdispersed energy modulation agents would provide a controlled anduniform way to heat a vat of material in a biological or chemicalprocess.

Photodeactivation

In many industrial processes, especially food and beverage industries,yeasts are used to produce changes in a medium such as the conversion ofsugars in the raw product. One particularly prominent example is in thewine industry. Stopping the wine from felinenting any further wouldpreserve the current level of sweetness. Likewise, allowing the wine tocontinue fermenting further would only make the wine less sweet witheach passing day. Eventually the wine would become completely dry atwhich time the fermentation would stop on its own. This is becauseduring the fermentation process yeast turns the sugar into alcohol.

Wanting to stop a fermentation is all good in and of itself. Butunfortunately, there is really no practical way to successfully stop afermentation dead in its tracks. Additives such as sulphite and sorbatecan be added to stabilize a fermented product and stop additionalfeintentation. Many winemakers will turn to sulfites such as that foundin Sodium Bisulfite or Campden tablets for the answer. But, these twoitems are not capable of reliably killing enough of the yeast toguarantee a complete stop of the activity—at least not at normal dosesthat leave the wine still drinkable.

Once the bulk of the sulfites from either of these ingredients dissipatefrom the wine into the air—as sulfites do—there is a very strong chancethat the remaining few live yeast cells will start multiplying andfetnienting again if given enough time. This usually happens at a mostinconvenient time, like after the wine has been bottled and stowed away.

Potassium sorbate is another ingredient that many winemakers considerwhen trying to stop a wine from fermenting any further. There is a lotof misunderstanding surrounding this product. It is typically called forby home wine making books when sweetening a wine. This is a situationwhere the fermentation has already completed and is ready for bottling.One adds the potassium sorbate along with the sugar that is added forsweetening.

The potassium sorbate stops the yeast from fermenting the newly addedsugar. So, many winemakers assume potassium sorbate can stop an activefelmentation as well, but, potassium sorbate does not kill the yeast atall, but rather it makes the yeast sterile. In other words, it impairsthe yeast's ability to reproduce itself. But, it does not hinder theyeast's ability to ferment sugar into alcohol.

Ultraviolet light is known to destroy yeast cultures, but has restrictedapplications due to the inability of UV light to penetrate throughoutthe fluid medium. While heat can be used to destroy the yeast activity,cooking of the product may be premature or may produce undesirablechanges in the consistency and taste. For liquid or fluid food products,the same techniques described above for liquid pasteurization could beused for the invention described here. For non-liquid products, energymodulation agents with little and preferably no toxicity (e.g. Fe oxidesor titanium oxides) could be added. Here, the concentration of theseadditives would likely be limited by any unexpected changes in taste.

Photoactivated Cross-linking and Curing of Polymers

In this application, luminescing particles (or energy modulation agents)are provided and distributed into an uncured polymer based medium forthe activation of photosensitive agents in the medium to promotecross-linking and curing of the polymer based medium. Additionally, theplasmonics agents can be included to enhance the effect of the incidentradiation or the internally generated radiation. In one embodiment, theplasmonics agents are complexed with the luminescent particles or otherenergy modulation agents prior to being added to the polymer.

As noted above, for adhesive and surface coating applications, lightactivated processing is limited due to the penetration depth of UV lightinto the processed medium. In light activated adhesive and surfacecoating processing, the primary limitation is that the material to becured must see the light—both in type (wavelength or spectraldistribution) and intensity. This limitation has meant that one mediumtypically has to transmit the appropriate light. In adhesive and surfacecoating applications, any “shaded” area will require a secondary curemechanism, increasing cure time over the non-shaded areas and furtherdelaying cure time due to the existent of a sealed skin through whichsubsequent curing must proceed.

Conventionally, moisture-curing mechanisms, heat-curing mechanisms, andphoto-initiated curing mechanisms are used to initiate cure, i.e.,cross-linking, of reactive compositions, such as reactive silicones,polymers, and adhesives. These mechanisms are based on eithercondensation reactions, whereby moisture hydrolyzes certain groups, oraddition reactions that can be initiated by a form of energy, such aselectromagnetic radiation or heat.

The invention described herein can use any of the following lightactivated curing polymers as well as others known in the art to whichthe luminescing particles (or energy modulation agents) are added.

For example, one suitable light activated polymer compound includes UVcuring silicones having methacrylate functional groups. U.S. Pat. No.4,675,346 to Lin, the disclosure of which is hereby expresslyincorporated herein by reference, is directed to UV curable siliconecompositions including at least 50% of a specific type of siliconeresin, at least 10% of a fumed silica filler and a photoinitiator, andcured compositions thereof. Other known UV curing silicone compositionssuitable for the invention include organopolysiloxane containing a(meth)acrylate functional group, a photosensitizer, and a solvent, whichcures to a hard film. Other known UV curing silicone compositionssuitable for the invention include compositions of an organopolysiloxanehaving an average of at least one acryloxy and/or methacryloxy group permolecule; a low molecular weight polyacrylyl crosslinking agent; and aphotosensitizer.

Loctite Corporation has designed and developed UV and UV/moisture dualcurable silicone compositions, which also demonstrate high resistance toflammability and combustibility, where the flame-retardant component isa combination of hydrated alumina and a member selected from the groupconsisting of organo ligand complexes of transition metals,organosiloxane ligand complexes of transition metals, and combinationsthereof. See U.S. Pat. Nos. 6,281,261 and 6,323,253 to Bennington. Theseformulations are also suitable for the invention.

Other known UV photoactivatable silicones include siliconesfunctionalized with for example carboxylate, maleate, cinnamate andcombinations thereof. These formulations are also suitable for theinvention. Other known UV photoactivatable silicones suitable for theinvention include benzoin ethers (“UV free radical generator”) and afree-radical polymerizable functional silicone polymers, as described inU.S. Pat. No. 6,051,625 whose content is incorporated herein byreference in its entirety. The UV free radical generator (i.e., thebenzoin ether) is contained at from 0.001 to 10 wt % based on the totalweight of the curable composition. Free radicals produced by irradiatingthe composition function as initiators of the polymerization reaction,and the free radical generator can be added in a catalytic quantityrelative to the polymerizable functionality in the subject composition.Further included in these silione resins can be silicon-bonded divalentoxygen atom compounds which can fonni a siloxane bond while theremaining oxygen in each case can be bonded to another silicon to form asiloxane bond, or can be bonded to methyl or ethyl to form an alkoxygroup, or can be bonded to hydrogen to form silanol. Such compounds caninclude trimethylsilyl, dimethylsilyl, phenyldimethylsilyl,vinyldimethylsilyl, trifluoropropyldimethylsilyl,(4-vinylphenyl)dimethylsilyl, (vinylbenzyl)dimethylsilyl, and(vinylphenethyl)dimethylsilyl.

The photoinitiator component of the invention is not limited to thosefree radical generators given above, but may be any photoinitiator knownin the art, including the aforementioned benzoin and substitutedbenzoins (such as alkyl ester substituted benzoins), Michler's ketone,dialkoxyacetophenones, such as diethoxyacetophenone (“DEAP”),benzophenone and substituted benzophenones, acetophenone and substitutedacetophenones, and xanthone and substituted xanthones. Other desirablephotoinitiators include DEAP, benzoin methyl ether, benzoin ethyl ether,benzoin isopropyl ether, diethoxyxanthone, chloro-thio-xanthone,azo-bisisobutyronitrile, N-methyl diethanolaminebenzophenone, andmixtures thereof. Visible light initiators include camphoquinone,peroxyester initiators and non-fluorene-carboxylic acid peroxyesters.

Commercially available examples of photoinitiators suitable for theinvention include those from Vantico, Inc., Brewster, N.Y. under theIRGACURE and DAROCUR tradenames, specifically IRGACURE 184(1-hydroxycyclohexyl phenyl ketone), 907(2-methyl-1-[4-(methylthio)phenyl]-2-morpholino propan-1-one), 369(2-benzyl-2-N,N-dimethylamino-1-(4-morpholinophenyl)-1-butanone), 500(the combination of 1-hydroxy cyclohexyl phenyl ketone andbenzophenone), 651 (2,2-dimethoxy-2-phenyl acetophenone), 1700 (thecombination of bis(2,6-dimethoxybenzoyl-2,4,4-trimethyl pentyl)phosphine oxide and 2-hydroxy-2-methyl-1-phenyl-propan-1-one), and 819[bis(2,4,6-trimethyl benzoyl)phenyl phosphine oxide] and DAROCUR 1173(2-hydroxy-2-methyl-1-phenyl-1-propane) and 4265 (the combination of2,4,6-trimethylbenzoyldiphenyl-phosphine oxide and2-hydroxy-2-methyl-1-phenyl-propan-1-one); and IRGACURE 784DC(bis(.eta..sup.5-2,4-cyclopentadien-1-yl)-bis[2,6-difluoro-3-(1H-pyrrol-1--yl)phenyl]titanium)

Generally, the amount of photoinitiator (or free radical generators)should be in the range of about 0 1% to about 10% by weight, such asabout 2 to about 6% by weight. The free radical generator concentrationfor benzoin ether is generally from 0.01 to 5% based on the total weightof the curable composition.

A moisture cure catalyst can also be included in an amount effective tocure the composition. For example, from about 0.1 to about 5% by weight,such as about 0.25 to about 2.5% by weight, of the moisture curecatalyst can be used in the invention to facilitate the cure processbeyond that of photo-activated curing. Examples of such catalystsinclude organic compounds of titanium, tin, zirconium and combinationsthereof. Tetraisopropoxytitanate and tetrabutoxytitanate are suitable asmoisture cure catalyst . See also U.S. Pat. No. 4,111,890, thedisclosure of which is expressly incorporated herein by reference.

Included in the conventional silicone composition (and other inorganicand organic adhesive polymers) suitable for the invention are variousinorganic fillers. For example, hollow microspheres supplied by Kishunder the trade name Q-CEL are free flowing powders, white in color.Generally, these borosilicate hollow microspheres are promoted asextenders in reactive resin systems, ordinarily to replace heavyfillers, such as calcium carbonate, thereby lowering the weight ofcomposite materials formed therewith. Q-CEL 5019 hollow microspheres areconstructed of a borosilicate, with a liquid displacement density of0.19 g/cm², a mean particle size of 70 microns, and a particle sizerange of 10-150 um. Other Q-CEL products are shown below in tabularfouu. Another commercially available hollow glass microsphere is sold byKish under the trade name SPHERICEL. SPHEREICEL 110P8 has a meanparticle size of about 11.7 microns, and a crush strength of greaterthan 10,000 psi. Yet other commercially available hollow glassmicrosphere are sold by the Schundler Company, Metuchen, N. J. under thePERLITE tradename, Whitehouse Scientific Ltd Chester, UK and 3M,Minneapolis, Minn. under the SCOTCHLTTE tradename.

In general, these inorganic filler components (and others such as fumedsilica) add structural properties to the cured composition, as well asconfers flowability properties to the composition in the uncured stateand increase the transmissivity for the UV cure radiation. When present,the fumed silica can be used at a level of up to about 50 weightpercent, with a range of about 4 to at least about 10 weight percent,being desirable. While the precise level of silica may vary depending onthe characteristics of the particular silica and the desired propertiesof the composition and the reaction product thereof, care should beexercised by those persons of ordinary skill in the art to allow for anappropriate level of transmissivity of the inventive compositions topermit a UV cure to occur.

Desirable hydrophobic silicas include hexamethyldisilazane-treatedsilicas, such as those commercially available from Wacker-Chemie,Adrian, Mich. under the trade designation HDK-2000. Others includepolydimethylsiloxane-treated silicas, such as those commerciallyavailable from Cabot Corporation under the trade designation CAB-O-SILN70-TS, or Degussa Corporation under the trade designation AEROSIL R202.Still other silicas include trialkoxyalkyl silane-treated silicas, suchas the trimethoxyoctyl silane-treated silica commercially available fromDegussa under the trade designation AEROSIL R805; and 3-dimethyldichlorosilane-treated silicas commercially available from Degussa underthe trade designation R972, R974 and R976.

While these inorganic fillers have extended the use of conventional UVcured silicone systems to pei nit the curing of materials beyond a skindepth of UV penetration, these inorganic fillers alone do not overcomeshadowing effects and suffer from UV scattering which effectively makesfor a smaller penetration depth. In the invention described herein, theinclusion of these inorganic fillers along with luminescing particlesprovide a mechanism by which uniform light activated cures can occurdeep inside of the body of adhesive-soliditied assemblies in regionsthat would normally be shadowed or not with the reach of external UV orother light sources.

Accordingly, in this example of the invention described herein,conventional silicone and polymeric adhesive or release or coatingcompositions are prepared using conventional mixing, heating, andincubation techniques. Included in these conventional compositions areluminescing particles. These luminescing particle containingcompositions can then be applied to surfaces of objects to be fixedtogether or to surfaces where a hard coating is desired or cast in acurable form for the production of molded objects. The luminescingparticles in these compositions upon activation will produce radiantlight for photoactivated cure of the luminescing particle containingpolymer composition. The density of luminescing particles in thesecompositions will depend on the “light transparency” of the luminescingparticle containing composition. Where these compositions contain asignificant amount of the inorganic filler as discussed above, theconcentration of luminescing particles can be reduced for example ascompared to a composition with a black color pigment where the lighttransparency will be significantly reduced.

One advantage of the invention described here as seen from this exampleis that now color pigments can be included in the light curable resinswithout significant compromise in the cured product performance. Thesecolor pigments may include one or more colored pigments well known tothose of ordinary skill in the art. Such pigments are generally metaloxides and include, but are not limited to, titanium dioxide, ironoxides, organic complexes, mica, talc and quartz. One pigment may beused, or a combination of two or more pigments may be utilized.Different colors can be obtained by choosing proper pigments andcombining them in a similar fashion as set forth in the followingexamples with the necessary adjustments, common in the paint industry,being made. Accordingly, in one embodiment of the invention, these colorpigments including carbon black may also be included as an opticallyopaque materials to limit the propagation of internally generated lightfrom the point of generation.

U.S. Pat. No. 7,294,656 to Bach et al., the entire disclosure of whichis incorporated herein by reference, describes a non-aqueous compositioncurable by UV radiation broadly containing a mixture of two UV curableurethane acrylates that have several advantages over conventionalradiation-curable compositions. The Bache et al. compositions can becured in a relatively short time using UV-C (200-280 nm), UV-B (280-320nm), UV-A (320-400 nm) and visible (400 nm and above) radiation. Inparticular, Bache et al compositions can be cured using radiation havinga wavelength of 320 nm or more. When fully cured (regardless of the typeof radiation used), the Bach et al. compositions exhibit hardnesses andimpact resistances at least comparable to conventional coatings.

In the invention described here, the luminescing particles (or energymodulation agents) described above are added to these Bach et al.compositions, optionally including in one embodiment various colorpigments. Due to the fact that the exterior energy source penetratesthroughout the entirety of the Bach et al. compositions, thicker surfacecoatings can be realized. Further, the coatings can be applied tointricate surfaces having for example been prepared with recesses orprotrusions. Curing with the recesses and around the protrusions withoutbeing limited by conventional UV shading will likely provide enhancedadherence of the surface coating to the work piece.

Moreover, in one embodiment of the invention, an external energy sourceof the initiation energy can be directed to a structural element inwhich a gap (or crack) therein was filled with an uncuredradiation-curable medium (such as those described above). The internallygenerated light will cure the uncured radiation-curable medium in thegap (or crack) thereby providing a repair to the structure beingirradiated.

Presently, there is available commercial epoxy systems which utilizeepoxy resin injection for the structural restoration of concrete. Epoxyinjection is very often the only alternative to complete replacement ofa structure. It therefore results in great cost savings. Besides fillingthe cracks, epoxy injection is known to protect rebar in the concreteand to stop water leakage. Commercially, the epoxy injection resinprovides a system for welding cracks which restores the originalstrength and loading originally designed into the concrete. Typically,low viscosity resins are pressure injected into the cracks. Often holesare drilled near or into the cracks to provide a conduit for pumping theresin into the cracks.

It however, takes time for the resin to penetrate into the thinner, evenhair line cracks. Unfortunately, time is limited in the presentcommercial systems due to the fact that the resins are premixed withhardeners whose time to cure sets an upper limit for how long the lowviscosity resin can flow into the cracks. Furtheintore, time to completerepair is an issue in many industrial repairs as the hardener is usuallypresent in a concentration high enough to have the resin set for examplein twenty four (24) hours. Moreover, with traditional resin methods, itis not possible to induce curing at specific regions of interest sinceall the areas of the resin will be cured

The present invention offers a number of advantages. Firstly, the resinof the present invention will be a photactivated resin which will notsubstantially cure until the x-ray source generates internal light toactivate the photoinitiators. This provides more flexibility in pumpingand waiting for complete crack fill. Secondly, once the photoactivatableresin is in place, its cure is then activated, and the cure occurs at arate not controlled by the convention hardening reaction. Thirdly, thex-ray penetration through the concrete and the crack region will providea more unifor,u mechanism for cure of the resins, with the deep cracksbeing as likely to fully cure as the narrow cracks which may extenddeeper into the material. Furthermore, the present invention allows thepossibility to cure only the specific areas of interest, i e where theX-ray is irradiated.

In another embodiment of the present invention, the external energysource can be a directed or focused beam of the initiation energy whichcures an uncured radiation-curable medium to produce a patternedelement. In this embodiment, the structure holding or at least partiallyenclosing the uncured radiation-curable medium can be a structure opaqueto visible light. In this manner, the uncured radiation-curable medium(which normally would be photoactivated upon exposure to ambient light)can be transported without premature curing. In this embodiment, thecuring would be activated for example by directed one or several focusedbeams of x-rays whose overlap generates regions in the structure holdingor at least partially enclosing the uncured radiation-curable mediumwhere the generated UV or visible light from the energy modulationagents in the medium would be of sufficient intensity to activate thephotoinitiators. In this manner, precise three-dimensional andtwo-dimensional patterning can be performed. In a similar embodiment,upconverting energy modulation agents could be used when the structureis transmissive of for example infrared or microwave frequencies. Theinitiation energy from, for example IR lasers, would be directed andfocused into the structure holding or at least partially enclosing theuncured radiation-curable medium.

As an example in another embodiment, a patterned element such as adevice (such as plug to close a specific internal hole or path ways) canbe fabricated (e.g., cured) inside structures (e.g., building materials,man-made or natural underground storage tank, internal organs of humanbody, etc) using energy excitation (e.g., X ray) from the outside ofsuch structures. Another application of this technique would involve thefabrication of orthopedic structures inside the body, where the curableresin was introduced locally at the point of the orthopedic structure tobe formed and a directed or focused x-ray beam cured the structure.

Accordingly, in another embodiment of the present invention, there isprovided a method (and associated system) for producing a patternedelement inside a structure. The method places inside the structure aradiation curable medium including at least one of a plasmonics agentand an energy modulation agent. The energy modulation agent isconfigured to emit light into the medium upon interaction with aninitiation energy. The method applies to the medium the initiationenergy from a directed or focused energy source. The applied initiationenergy interacts with the plasmonics agent or the energy modulationagent to generate light at local regions inside the structure to curelocally the radiation curable medium.

As noted above, this method can form for the patterned element a plug toclose a hole or pathway in the structure such as for example holes orpathways in a building material, a man-made or natural undergroundstorage tank, or an internal organ in a human or animal body. The methodcan form for the patterned element a prosthetic device at a local pointin the body of a human or animal.

The method can further localize the curing by placing in the radiationcurable medium optically dense materials (such as the color pigmentsdiscussed above) to reduce propagation of the generated light from thepoint of generation.

Computer-Assisted Control

In one embodiment of the invention, there is provided a computerimplemented system for designing and selecting suitable combinations ofinitiation energy source, energy modulation agent, and activatableagent. For example, the computer system 5 can include a centralprocessing unit (CPU) having a storage medium on which is provided: adatabase of excitable compounds, a first computation module for aphotoactivatable agent or energy transfer agent, and a secondcomputation module predicting the requisite energy flux needed tosufficiently activate the or energy transfer agent or photoactivatableagent.

FIG. 4 illustrates a computer system 1201 for implementing variousembodiments of the invention. The computer system 1201 may be used asthe computer system 5 to perform any or all of the functions describedabove. The computer system 1201 includes a bus 1202 or othercommunication mechanism for communicating information, and a processor1203 coupled with the bus 1202 for processing the information. Thecomputer system 1201 also includes a main memory 1204, such as a randomaccess memory (RAM) or other dynamic storage device (e.g., dynamic RAM(DRAM), static RAM (SRAM), and synchronous DRAM (SDRAM)), coupled to thebus 1202 for storing information and instructions to be executed byprocessor 1203. In addition, the main memory 1204 may be used forstoring temporary variables or other intermediate information during theexecution of instructions by the processor 1203. The computer system1201 further ncludes a read only memory (ROM) 1205 or other staticstorage device (e.g., programmable read only memory (PROM), erasablePROM (EPROM), and electrically erasable PROM (EEPROM)) coupled to thebus 1202 for storing static infoimation and instructions for theprocessor 1203.

The computer system 1201 also includes a disk controller 1206 coupled tothe bus 1202 to control one or more storage devices for storinginformation and instructions, such as a magnetic hard disk 1207, and aremovable media drive 1208 (e.g., floppy disk drive, read-only compactdisc drive, read/write compact disc drive, compact disc jukebox, tapedrive, and removable magneto-optical drive). The storage devices may beadded to the computer system 1201 using an appropriate device interface(e.g., small computer system interface (SCSI), integrated deviceelectronics (IDE), enhanced-IDE (E-IDE), direct memory access (DMA), orultra-DMA).

The computer system 1201 may also include special purpose logic devices(e.g., application specific integrated circuits (ASICs)) or configurablelogic devices (e.g., simple programmable logic devices (SPLDs), complexprogrammable logic devices (CPLDs), and field programmable gate arrays(FPGAs)).

The computer system 1201 may also include a display controller 1209coupled to the bus 1202 to control a display, such as a cathode ray tube(CRT), for displaying information to a computer user. The computersystem includes input devices, such as a keyboard and a pointing device,for interacting with a computer user and providing information to theprocessor 1203. The pointing device, for example, may be a mouse, atrackball, or a pointing stick for communicating direction informationand command selections to the processor 1203 and for controlling cursormovement on the display. In addition, a printer may provide printedlistings of data stored and/or generated by the computer system 1201.

The computer system 1201 performs a portion or all of the processingsteps of the invention (such as for example those described in relationto FIG. 5) in response to the processor 1203 executing one or moresequences of one or more instructions contained in a memory, such as themain memory 1204. Such instructions may be read into the main memory1204 from another computer readable medium, such as a hard disk 1207 ora removable media drive 1208. One or more processors in amulti-processing arrangement may also be employed to execute thesequences of instructions contained in main memory 1204. In alternativeembodiments, hard-wired circuitry may be used in place of or incombination with software instructions. Thus, embodiments are notlimited to any specific combination of hardware circuitry and software.

As stated above, the computer system 1201 includes at least one computerreadable medium or memory for holding instructions programmed accordingto the teachings of the invention and for containing data structures,tables, records, or other data described herein. Examples of computerreadable media are compact discs, hard disks, floppy disks, tape,magneto-optical disks, PROMs (EPROM, EEPROM, flash EPROM), DRAM, SRAM,SDRAM, or any other magnetic medium, compact discs (e.g CD-ROM), or anyother optical medium, punch cards, paper tape, or other physical mediumwith patterns of holes, a carrier wave (described below), or any othermedium from which a computer can read.

Stored on any one or on a combination of computer readable media, theinvention includes software for controlling the computer system 1201,for driving a device or devices for implementing the invention, and forenabling the computer system 1201 to interact with a human user. Suchsoftware may include, but is not limited to, device drivers, operatingsystems, development tools, and applications software. Such computerreadable media further includes the computer program product of theinvention for performing all or a portion (if processing is distributed)of the processing performed in implementing the invention.

The computer code devices of the invention may be any interpretable orexecutable code mechanism, including but not limited to scripts,interpretable programs, dynamic link libraries (DLLs), Java classes, andcomplete executable programs. Moreover, parts of the processing of theinvention may be distributed for better performance, reliability, and/orcost.

The term “computer readable medium” as used herein refers to any mediumthat participates in providing instructions to the processor 1203 forexecution. A computer readable medium may take many forms, including butnot limited to, non-volatile media, volatile media, and transmissionmedia. Non-volatile media includes, for example, optical, magneticdisks, and magneto-optical disks, such as the hard disk 1207 or theremovable media drive 1208. Volatile media includes dynamic memory, suchas the main memory 1204. Transmission media includes coaxial cables,copper wire and fiber optics, including the wires that make up the bus1202. Transmission media also may also take the form of acoustic orlight waves, such as those generated during radio wave and infrared datacommunications.

Various forms of computer readable media may be involved in carrying outone or more sequences of one or more instructions to processor 1203 forexecution. For example, the instructions may initially be carried on amagnetic disk of a remote computer. The remote computer can load theinstructions for implementing all or a portion of the invention remotelyinto a dynamic memory and send the instructions over a telephone lineusing a modem. A modem local to the computer system 1201 may receive thedata on the telephone line and use an infrared transmitter to convertthe data to an infrared signal. An infrared detector coupled to the bus1202 can receive the data carried in the infrared signal and place thedata on the bus 1202. The bus 1202 carries the data to the main memory1204, from which the processor 1203 retrieves and executes theinstructions. The instructions received by the main memory 1204 mayoptionally be stored on storage device 1207 or 1208 either before orafter execution by processor 1203.

The computer system 1201 also includes a communication interface 1213coupled to the bus 1202. The communication interface 1213 provides atwo-way data communication coupling to a network link 1214 that isconnected to, for example, a local area network (LAN) 1215, or toanother communications network 1216 such as the Internet. For example,the communication interface 1213 may be a network interface card toattach to any packet switched LAN. As another example, the communicationinterface 1213 may be an asymmetrical digital subscriber line (ADSL)card, an integrated services digital network (ISDN) card or a modem toprovide a data communication connection to a corresponding type ofcommunications line. Wireless links may also be implemented. In any suchimplementation, the communication interface 1213 sends and receiveselectrical, electromagnetic or optical signals that carry digital datastreams representing various types of information.

The network link 1214 typically provides data communication through oneor more networks to other data devices. For example, the network link1214 may provide a connection to another computer through a localnetwork 1215 (e.g., a LAN) or through equipment operated by a serviceprovider, which provides communication services through a communicationsnetwork 1216. The local network 1214 and the communications network 1216use, for example, electrical, electromagnetic, or optical signals thatcarry digital data streams, and the associated physical layer (e.g., CAT5 cable, coaxial cable, optical fiber, etc). The signals through thevarious networks and the signals on the network link 1214 and throughthe communication interface 1213, which carry the digital data to andfrom the computer system 1201 maybe implemented in baseband signals, orcarrier wave based signals. The baseband signals convey the digital dataas unmodulated electrical pulses that are descriptive of a stream ofdigital data bits, where the tei m “bits” is to be construed broadly tomean symbol, where each symbol conveys at least one or more informationbits. The digital data may also be used to modulate a carrier wave, suchas with amplitude, phase and/or frequency shift keyed signals that arepropagated over a conductive media, or transmitted as electromagneticwaves through a propagation medium. Thus, the digital data may be sentas unmodulated baseband data through a “wired” communication channeland/or sent within a predetermined frequency band, different thanbaseband, by modulating a carrier wave. The computer system 1201 cantransmit and receive data, including program code, through thenetwork(s) 1215 and 1216, the network link 1214, and the communicationinterface 1213. Moreover, the network link 1214 may provide a connectionthrough a LAN 1215 to a mobile device 1217 such as a personal digitalassistant (PDA) laptop computer, or cellular telephone.

The exemplary energy spectrum previously noted in FIG. 1 may also beused in this computer-implemented system.

The reagents and chemicals useful for methods and systems of theinvention may be packaged in kits to facilitate application of theinvention. In one exemplary embodiment, a kit would comprise at leastone activatable agent capable of producing a predetermined cellularchange, at least one energy modulation agent capable of activating theat least one activatable agent when energized, at least one plasmonicsagent that can enhance applied initiation energy such that the enhancedinitiation energy activates the at least one activatable agent whichproduces a change in the medium when activated. and containers suitablefor storing the agents in stable form, and further comprisinginstructions for administering the at least one activatable agent and atleast one energy modulation agent to a medium, and for applying aninitiation energy from an initiation energy source to activate theactivatable agent. The instructions could be in any desired form,including but not limited to, printed on a kit insert, printed on one ormore containers, as well as electronically stored instructions providedon an electronic storage medium, such as a computer readable storagemedium. Also optionally included is a software package on a computerreadable storage medium that permits the user to integrate the informiiation and calculate a control dose, to calculate and control intensityof the irradiation source.

System Implementation

In one embodiment of the invention, there is provided a first system forproducing a change in a medium disposed in an artificial container. Thefirst system includes a mechanism configured to supply in the medium atleast one of a plasmonics agent and an activatable agent. The plasmonicsagent enhances or modifies energy in a vicinity of itself. In oneexample, the plasmonics agent enhances or modifies the appliedinitiation energy such that the enhanced initiation energy producesdirectly or indirectly the change in the medium. The system includes aninitiation energy source configured to apply an initiation energythrough the artificial container to the medium to activate the at leastone activatable agent in the medium.

In one embodiment, the energy modulation agent converts the appliedinitiation energy and produces light at an energy different from theapplied initiation energy. The plasmonics agent can enhance the lightfrom the at least one energy modulation agent. In one embodiment, theapplied initiation energy source is an external initiation energysource. In one embodiment, the applied initiation energy source is asource that is at least partially in a container holding the medium.

The medium in one embodiment is substantially transparent to theinitiation energy. For example, if the medium s a liquid or fluid foodproduct such as orange juice which has a substantial amount of suspendedsolids, then UV light for example as described above and even visiblelight will be substantially absorbed and/or scattered by the orangejuice medium. Furthermore, microwave energy will likewise be absorbed bythis medium. However, an initiation energy source such as an X-raysource will essentially transmit entirely through for example an orangejuice medium. The effect is the medium can now be totally illuminatedwith the external initiation energy source.

Other sources and tuned to specific wavelengths may also be used as theinitiation energy source. These sources would take advantage of an“optical window” in the medium where for example a particular wavelengthof light would not be absorbed. Water selectively scatters and absorbscertain wavelengths of visible light. The long wavelengths of the lightspectrum—red, yellow, and orange—can penetrate to approximately 15, 30,and 50 meters (49, 98, and 164 feet), respectively, while the shortwavelengths of the light spectrum—violet, blue and green—can penetratefurther. Thus, for many aqueous based systems, non-high energy X-raysources may not be needed. In those situations, energy modulation agentsand plasmonics agents would be added whose interaction with the incidentlight would produce for example photoactivation of catalysts in theaqueous medium. Light produced from the energy modulation agent can alsobe enhanced by the plasmonics agents in the medium.

Accordingly, depending on the medium and the energy modulation agent andthe activatable agent, the initiation energy source can include at leastone of an X-ray source, a gamma ray source, an electron beam source, anUV radiation source, a visible and infrared source, a microwave source,or a radio wave source. The initiation energy source can then be anenergy source emitting one of electromagnetic energy, acoustic energy,or theiinal energy. The initiation energy source can then be an energysource emitting a wavelength whose depth of penetration penetratesthroughout the medium. The initiation energy in one embodiment may bescattered or absorbed in the medium, but the plasmonics agents makeuseful the remnant light. The medium to be effected can be a medium tobe fermented, sterilized, or cold pasteurized. The medium to be effectedcan include bacteria, viruses, yeasts, and fungi.

The activatable agents can be photoactivatable agents such as thephotocages (described elsewhere) such that upon exposure to theinitiation energy source, the photocage disassociates rendering anactive agent available. The activatable agents can include agents suchas psoralens, pyrene cholesteryloleate, acridine, porphyrin,fluorescein, rhodamine, 16-diazorcortisone, ethidium, transition metalcomplexes of bleomycin, transition metal complexes of deglycobleomycinorganoplatinum complexes, alloxazines, vitamin Ks, vitamin L, vitaminmetabolites, vitamin precursors, naphthoquinones, naphthalenes,naphthols and derivatives thereof having planar molecularconforminations, porphorinporphyrins, dyes and phenothiazinederivatives, coumarins, quinolones, quinones, and anthroquinones. Theactivatable agents can include photocatalysts such as TiO₂, ZnO, CdS,CdSe, SnO₂, SrTiO₃, WO₃, Fe₂O₃, and Ta₂O₅ particles.

The first system can include a mechanism configured to provide in themedium at least one energy modulation agent which converts theinitiation energy to an activation energy for activation of theactivatable agent(s). The energy modulation agent(s) can be a photonemitter such a phosphorescent compounds, chemiluminescent compounds, andbioluminescent compounds. The energy modulation agent(s) can be upconversion or down conversion agents. The energy modulation agent(s) canbe luminescent particles which emit light upon exposure to saidinitiation energy. The energy modulation agent(s) can be nanotubes,nanoparticles, chemilumiscent particles, and bioluminescent particles,and mixtures thereof. The luminescent particles can be nanoparticles ofsemiconducting or metallic materials. The luminescent particles can bechemiluminescent particles which show enhanced chemiluminescence uponexposure to microwaves.

The first system can include a mechanism configured to provide in themedium plasmonics-agents including metal nanostructures such as forexample nanospheres, nanorods, nanocubes, nanopyramids, nanoshells,multi-layer nanoshells, and combinations thereof. The form and structureof these plasmonics-agents can include the probe structures detailedabove.

Depending on the initiation energy source, the system can include acontainer for the medium that is permeable to the applied initiationenergy. For example, for an X-ray source, the container can be made ofaluminum, quartz, glass, or plastic. For a microwave source, thecontainer can be made of quartz, glass, or plastic. Furthei more, thecontainer can be a container which receives and transmits the initiationenergy to fluid products to pasteurize the fluid products, or can be acontainer which receives and transmits the initiation energy to fluidproducts to remediate contaminants in the fluid products.

In another embodiment of the invention, there is provided a secondsystem for curing a radiation-curable medium. The second system includesa mechanism configured to supply an uncured radiation-curable mediumincluding at least one plasmonics agent and at least one activatableagent which produces a change in the radiation-curable medium whenactivated, and further includes an applied initiation energy sourceconfigured to apply initiation energy to a composition including theuncured radiation-curable medium, the plasmonics agent, and the energymodulation agent. The energy modulation agent as described above absorbsthe initiation energy and converts the initiation energy to anactivation energy capable of curing the uncured medium (i.e., promotingpolymerization of polymers in the uncured medium). The plasmonics agentenhances the applied initiation energy such that the enhanced initiationenergy directly or indirectly cures the medium by polymerization ofpolymers in the medium. For example, the plasmonics agent can enhancethe activation energy light such that enhanced light activates the atleast one photoactivatable agent to polymerize polymers in the medium.In another example, activation of the energy modulation gaent produces alight which activates the at least one photoactivatable agent topolymerize polymers in the medium.

The second system has attributes similar to the first system describedabove and can further permit the at least one activatable agent toinclude a photoinitiator such as one of benzoin, substituted benzoins,alkyl ester substituted benzoins, Michler's ketone,dialkoxyacetophenones, diethoxyacetophenone, benzophenone, substitutedbenzophenones, acetophenone, substituted acetophenones, xanthone,substituted xanthenes, benzoin methyl ether, benzoin ethyl ether,benzoin isopropyl ether, di ethoxyxanthone, chloro-thio-xanthone,azo-bisisobutyronitrile, N-methyl diethanolaminebenzophenone,camphoquinone, peroxyester initiators, non-fluorene-carboxylic acidperoxyesters and mixtures thereof.

The second system can also include a mechanism configured to provide inthe medium plasmonics-agents including metal nanostructures such as forexample nanospheres, nanorods, nanocubes, nanopyramids, nanoshells,multi-layer nanoshells, and combinations thereof. The formin andstructure of these plasmonics-agents can include the probe structuresdetailed above.

The second system can include a container for the uncuredradiation-curable medium that is permeable to the applied initiationenergy. The container can be configured to contain the uncuredradiation-curable medium or to hold a mold of the uncuredradiation-curable medium. The container as before can be an aluminumcontainer, a quartz container, a glass container, or a plasticcontainer, depending on the applied initiation energy.

In one embodiment, an energy source (e.g., an external energy source) isconfigured to irradiate the uncured radiation-curable medium in a jointregion (or regions) adhering one region of a utensil to another regionof the utensil. In another embodiment, the energy source is configuredto irradiate the joint regions and thereby induce sterilization of thejoint regions due to the production of internal UV light inside thejoint regions. In another embodiment, the energy source is configured toirradiate a surface coating. In another embodiment, the energy source isconfigured to irradiate a mold of the radiation-curable medium.

The radiation-curable medium in the surface coating or in the mold or inother medium can include color pigments to add color to a finished curedproduct. The radiation-curable medium in the surface coating or in themold or in another medium can include fumed silica to promote strengthand enhance distribution of the internally generated light. Theradiation-curable medium in the surface coating or in the mold or inanother medium can include a moisture cure promoter to supplement thecure.

The second system provides one mechanism for production of novelradiation-cured articles, which include a radiation-cured medium, atleast one plasmonics agent, and at least one energy modulation agentdistributed throughout the medium. The energy modulation agent being asubstance which is capable of converting an applied energy to lightcapable of producing a cure for the radiation-cured medium. Theplasmonics agent enhances the applied initiation energy such that theenhanced initiation energy activates the energy modulation agents. Lightproduced from the energy modulation agent can also be enhanced by theplasmonics agents in the medium. The article can include luminescentparticles such as for example nanotubes, nanoparticles, chemilumi scentparticles, and bioluminescent particles, and mixtures thereof. Thearticle can include nanoparticles of semiconducting or metallicmaterials. The article can include chemiluminescent particles. Thearticle can include color pigments or fumed silica. The article caninclude plasmonics-agents including metal nanostructures such as forexample nanospheres, nanorods, nanocubes, nanopyramids, nanoshells,multi-layer nanoshells, and combinations thereof. The form and structureof these plasmonics-agents can include the probe structures detailedabove.

In another embodiment of the invention, there is provided a third systemfor producing a change in a medium disposed in an artificial container.The third system includes a mechanism configured to provide to themedium 1) an activatable agent and 2) at least one of a plasmonics agentand an energy modulation agent. The energy modulation agent converts aninitiation energy to an activation energy which then activates the atleast one activatable agent. The third system further includes anapplied initiation energy source configured to apply the initiationenergy through the artificial container to activate the at least oneactivatable agent in the medium. The plasmonics agent enhances ormodifies an energy in a vicinity of itself. In one example, theplasmonics agent enhances or modifies the applied initiation energy suchthat the enhanced initiation energy produces directly or indirectly thechange in the medium.

The third system has similar attributes to the first and second systemsdescribed above, and further includes encapsulated structures includingat least one of the energy modulation agent and the plasmonics agent.The encapsulated structures can include nanoparticles of the energymodulation agent encapsulated with a passivation layer or can includesealed quartz or glass tubes having the energy modulation agent inside.The encapsulated structures can include sealed tubes having theplasmonics agent disposed on an outside of the sealed tube (which may ormay not be exposed directly to the medium).

In another embodiment of the invention, there is provided a fourthsystem for producing a photo-stimulated change in a medium disposed inan artificial container. The fourth system includes a mechanismconfigured to provide in the medium at least one of a plasmonics agentand an energy modulation agent. The energy modulation agent converts aninitiation energy to an activation energy which then produces thephoto-stimulated change. The fourth system further includes aninitiation energy source configured to apply the initiation energy tothe medium to activate the at least one energy modulation agent in themedium. The plasmonics agent enhances or modifies an energy in avicinity of itself. In one example, the plasmonics agent enhances ormodifies the applied initiation energy such that the enhanced initiationenergy produces directly or indirectly the change in the medium. Thesystem can include encapsulated structures including therein the energymodulation agent. The encapsulated structures can include nanoparticlesof the energy modulation agent encapsulated with a passivation layer.The encapsulated structures can include sealed tubes having theplasmonics agent disposed on an outside of the sealed tube (which may ormay not be exposed directly to the medium).

The fourth system can include a container which receives and transmitsthe initiation energy to products within the medium. The products caninclude plastics, where the activation energy alters the surfacestructure of the plastics. The products can include polylactic acid(PLA) plastics and polyhydroxyalkanoates (PHA) plastics. In thisembodiment, the activation energy can photo-graft a molecular speciesonto a surface of the plastics.

Sterilization Methods and System Components

Optical techniques have been often used in sterilization procedures torender unwanted or harmful waterborne microorganisms incapable ofreproducing using ultraviolet light (specifically the spectral area ofUV-C, 200 to 280 nm range). Ultraviolet light in the UV-C is consideredthe most lethal range as a germicidal disinfectant (capable of alteringa living microorganism's DNA, and keeping the microorganism fromreproducing). UV-C, with 264 nanometers being the peak germicidalwavelength, is known as the germicidal spectrum. Although the UV-Cmethod is simple and effective, it is not particularly effective insamples (gas, liquids, particulates) enclosed on containers which do nottransmit UV light. The present invention provides techniques and systemsthat can use externally applied radiation such as X-ray forsterilization. While illustrated below with respect to X-rayirradiation, and as discussed above, other suitable fog ms of energycould be used provided the containers and medium to be sterilized wassufficiently transparent for the medium to be thoroughly irradiated.Examples of alternative sources and materials for upconvertingluminescence to higher energies have been discussed above.

FIGS. 27-44 show various embodiments of sterilization systems and probesthat can be used with X ray excitation. These systems are applicable ina number of the applications discussed above and as well as in othersterilization areas. The systems could thus be used in the waste waterdetoxification, blood sterilization, cold pasteurization, andphotodeactivation commercial applications discussed in the sectionsabove. These systems (like FIGS. 3B-3D) show the use of artificialcontainers in which the medium to be treated is disposed.

FIG. 27 shows one embodiment of a sterilization system of the inventionthat includes: a container and a material containing an X-ray energyconverter. The container holds a sample to be sterilized (e.g., liquid,gas, or particulates). X-ray radiation, capable of penetrating thecontainer wall, excites the material containing the X-ray excitationenergy converter (EEC), which is configured to emit emission light. TheEEC material is selected such that the emitted or luminescence lightoccurs in a spectral region that can be used for sterilization (e.g.,the ultraviolet spectral range).

FIG. 28 shows one embodiment of another sterilization system of theinvention that utilizes plasmonics and includes: a container, a materialcontaining an X-ray energy converter, a dielectric layer (e.g., silica),and a metal nanostructure (e.g., Au, Ag). The container holds a sampleto be sterilized (e.g., liquid, gas, or particulates). X-ray radiation,capable of penetrating the container wall, excites the materialcontaining the X-ray excitation energy converter (EEC), which in turnemits emission light. The EEC material is selected such that the emittedor luminescence light occurs in a spectral region that can be used forsterilization (e.g., an ultraviolet spectral range). The metalnanostructure is designed to amplify the luminescence light due to theplasmonics enhancement effect discussed above. The dielectric layer isdesigned to separate the material of the X-ray energy converter from themetal nanostructure in order to minimize or prevent possible quenchingof the luminescence. The optimal thickness of the dielectric layer isabout 1 to 5 nm such that the dielectric layer does not significantlyalter the plasmonics effect.

FIG. 29 shows another embodiment of a sterilization system of theinvention that includes: a container, a material containing an X-rayenergy converter, and a photo-active (PA) material. X-ray radiation,capable of penetrating the container wall, excites the materialcontaining the X-ray excitation energy converter (EEC), which in turnemits luminescence light. The EEC material is selected such that theemitted or luminescence light occurs in the spectral region that can beused to further excite the photo-active (PA) material. The photo-activematerial can be used for sterilization-purpose emission light (e.g.,luminescence) following excitation by the EEC luminescence light.Alternatively the PA material is replaced by or is a material that hasthe property of up/down energy conversion of the EEC emission light inorder to produce radiation at appropriate wavelengths for sterilizationpurposes (e.g., UV light to kill bacteria).

FIG. 30 shows another embodiment of a sterilization system of theinvention that includes: a container, a material including an X-rayenergy converter, a dielectric layer (e.g., silica), a metalnanostructure (e.g., Au, Ag), and a photo-active (PA) material. X-rayradiation, capable of penetrating the container wall, excites thematerial containing the X-ray excitation energy converter (EEC), whichin turn emits emission light. The EEC material is selected such that theemitted or luminescence light occurs in a spectral region that can beused to further excite a photo-active (PA) material. The photo-activematerial can be used for sterilization purpose e ss on light (e.g.,luminescence) following excitation by the EEC luminescence light.Alternatively the PA material is replaced by or is a material that hasthe property of up/down energy conversion of the EEC emission light inorder to produce radiation at appropriate wavelengths for sterilizationpurposes (e.g., UV light to kill bacteria). The metal nanostructure inthis embodiment is designed to amplify the luminescence light due to theplasmonics enhancement effect. The dielectric layer is designed toseparate the material containing X-ray energy converter and the metalnanostructure in order to prevent or minimize possible quenching of theluminescence.

FIG. 31 shows another embodiment of a sterilization system of theinvention that includes: a container and a material including an X-rayenergy converter with embedded metal nanoparticles included as part ofthe walls of the container. The container holds a sample to besterilized which can be a liquid, a gas, or particulates. The X-rayradiation, capable of penetrating the container wall, excites thematerial containing the X-ray excitation energy converter (EEC), whichin turn emits emission light. The EEC material is selected such that theemitted or luminescence light occurs in a spectral region that can beused for sterilization (e.g., the ultraviolet spectral range). In thisembodiment, the EEC material is contained in a matrix that also hasmetallic nanoparticles (1-100 nm diameter). The metallic nanoparticlesserve as plasmonics-active systems that are designed to enhance the EECemission light.

FIG. 32 shows another embodiment of a sterilization system of theinvention that includes: a container and a material including an X-rayenergy converter with embedded metal nanoparticles included as part ofthe walls of the container and included on re-entrant structures. Thisembodiment is designed such that a sample flow can have maximum contactwith the walls (including the re-entrant structures) of thesterilization system. The sample flowing through the container can beliquid, gas, or particulates. X-ray radiation, capable of penetratingthe container wall, excites the material containing the X-ray excitationenergy converter (EEC), which in turn emits emission light. The EECmaterial is selected such that the emitted or luminescence light occursin a spectral region that can be used for sterilization (e.g., theultraviolet spectral range). In this embodiment, the EEC material iscontained in a matrix that also has metallic nanoparticles (1-100 nmdiameter). The metallic nanoparticles serve has plasmonics-activesystems that are designed to enhance the EEC emission light.

FIG. 33 shows another embodiment of a sterilization system of theinvention that includes: a container, a material including an X-rayenergy converter, and a photo-active material. The container holds asample to be sterilized which can be a liquid, a gas, or particulates.X-ray radiation, capable of penetrating the container wall, excites thematerial containing X-ray excitation energy converter (EEC), which inturn emits emission light. The EEC material is selected such that theemitted or luminescence light occurs in the spectral region that can beused to further excite a photo-active (PA) material. The photo-activematerial can be used for sterilization purpose emission light (e.g.,luminescence) following excitation by the EEC luminescence light.Alternatively the PA material is replaced by or is a material that hasthe property of up/down energy conversion of the EEC emission light inorder to produce radiation at appropriate wavelengths for sterilizationpurposes (e.g., UV light to kill bacteria). In this embodiment, the PAmaterial (or up/down energy conversion material) is contained in amatrix that also has metallic nanoparticles (1-100 nm diameter). Themetallic nanoparticles serve has plasmonics-active systems that aredesigned to enhance the emission light.

FIG. 34 shows another embodiment of a sterilization system of theinvention that includes: a container, a material including an X-rayenergy converter with embedded metal nanoparticles included on an insidelayer on the walls of the container and included on re-entrantstructures, and a photo-active material. The container holds a sample tobe sterilized which can be a liquid, a gas, or particulates. Thisembodiment is designed such that a sample flow can have frequent contactwith the walls of the sterilization system. X-ray radiation, capable ofpenetrating the container wall, excites the material containing X-rayexcitation energy converter (EEC), which in turn emits emission light.The EEC material is selected such that the emitted or luminescence lightoccurs in the spectral region that can be used to further excite aphoto-active (PA) material. The photo-active material can be used forsterilization-purpose emission light (e.g., luminescence) followingexcitation by the EEC luminescence light. Alternatively, the PA materialis replaced by or is a material that has the property of up/down energyconversion of the EEC emission light in order to produce radiation atappropriate wavelengths for sterilization purposes (e.g., UV light tokill bacteria). In this embodiment, the PA material (or up/down energyconversion material) is contained in a matrix that also has metallicnanoparticles (1-100 nm diameter). The metallic nanoparticles serve hasplasmonics-active systems that are designed to enhance the emissionlight.

FIG. 35 shows another embodiment of a sterilization system of theinvention that includes: a container, a material including an X-rayenergy converter, and chemical receptors or bioreceptors used to capturetargets. The container holds a sample to be sterilized which can be aliquid, a gas, or particulates. X-ray radiation, capable of penetratingthe container wall, excites the material containing X-ray excitationenergy converter (EEC), which in turn emits emission light. The EECmaterial is selected such that the emitted or luminescence light occursin a spectral region that can be used for sterilization (e.g. theultraviolet spectral range). The layer of chemical receptors (e.g.,ligands specific to chemical groups) or bioreceptors (e.g., antibodies,surface cell receptors) is used to capture biochemical targets ofinterest. In this embodiment, the specific target compounds areselectively bound to the surface and are more effectively irradiated bythe emission light.

FIG. 36 shows another embodiment of a sterilization system of theinvention that includes: a container, a material including an X-rayenergy converter, a dielectric layer (e.g., silica), metalnanostructures (e.g., Au, Ag), and chemical receptors or bioreceptorsused to capture targets. The sample inside the container can be liquid,gas, or particulates. X-ray radiation, capable of penetrating thecontainer wall, excites the material containing X-ray excitation energyconverter (EEC), which in turn emits emission light. The EEC material isselected such that the emitted or luminescence light occurs in aspectral region that can be used for sterilization (e.g., theultraviolet spectral range). The metal nanostructure is designed toamplify the luminescence (or emitted) light due to the plasmonicsenhancement effect. The dielectric layer is designed to separate thematerial containing X-ray energy converter and the metal nanostructurein order to prevent or minimize possible quenching of the luminescence.The optimal thickness of the dielectric layer is about 1 to 5 nm suchthat the dielectric layer does not significantly affect the plasmonicseffect. The layer of chemical receptors (e.g., ligands specific tochemical groups) or bioreceptors (e.g., antibodies, surface cellreceptors) is used to capture biochemical targets of interest. In thisembodiment, the specific target compounds are selectively bound to thesurface and are more effectively irradiated by the emission light.

FIG. 37 shows another embodiment of a sterilization system of theinvention that includes: a container, a material including an X-rayenergy converter, a photo-active (PA) material, and chemical receptorsor bioreceptors used to capture targets. X-ray radiation, capable ofpenetrating the container wall, excites the material containing X-rayexcitation energy converter (EEC), which in turn emits luminescencelight. The EEC material is selected such that the emitted orluminescence light occurs in a spectral region that can be used tofurther excite a photo-active (PA) material. The photo-active materialcan be used for sterilization-purpose emission light (e.g.,luminescence) following excitation by the EEC luminescence light. Thelayer of chemical receptors (e.g., ligands specific to chemical groups)or bioreceptors (e.g., antibodies, surface cell receptors) is used tocapture biochemical targets of interest. In this embodiment, thespecific target compounds are selectively bound to the surface and aremore effectively irradiated by the emission light. Alternatively, the PAmaterial is replaced by or is a material that has the property ofup/down energy conversion of the EEC emission light in order to produceradiation at appropriate wavelengths for sterilization purposes (e.g.,UV light to kill bacteria).

FIG. 38 shows another embodiment of a sterilization system of theinvention that includes: a container, a material including an X-rayenergy converter, a photo-active (PA) material, a metal nanostructure(e.g., Au, Ag), a dielectric layer (e.g., silica), and chemicalreceptors or bioreceptors used to capture targets. X-ray radiation,capable of penetrating the container wall, excites the materialcontaining X-ray excitation energy converter (EEC), which in turn emitsemission light. The EEC material is selected such that the emitted orluminescence light occurs in a spectral region that can be used tofurther excite a photo-active (PA) material. The photo-active materialcan be used for sterilization-purpose emission light (e.g.,luminescence) following excitation by the EEC luminescence light.Alternatively, the PA material is replaced by or is a material that hasthe property of up/down energy conversion of the EEC emission light inorder to produce radiation at appropriate wavelengths for sterilizationpurposes (e.g UV light to kill bacteria). The metal nanostructure isdesigned to amplify the luminescence light due to the plasmonicsenhancement effect. The dielectric layer is designed to separate thematerial containing X-ray energy converter and the metal manostructurein order to prevent or minimize possible quenching of the luminescence.The layer of chemical receptors (e.g., ligands specific to chemicalgroups) or bioreceptors (e.g., antibodies, surface cell receptors) isused to capture biochemical targets of interest. In this embodiment, thespecific target compounds are selectively bound to the surface and aremore effectively irradiated by the emission light.

The invention can use these chemical receptors and bioreceptors oninterior walls contacting the medium to be sterilized in the othersystems shown herein.

FIG. 39 shows an embodiment of a sterilization probe system of theinvention that includes a container which can hold the medium to besterilized and a probe made of material containing an X-ray energyconverter. The sample inside the container can be liquid, gas, orparticulates. X-ray radiation, capable of penetrating the containerwall, excites the probe having the material containing X-ray excitationenergy converter (EEC), which in turn emits emission light. The EECmaterial is selected such that the emitted or luminescence light occursin a spectral region that can be used for sterilization (e.g., theultraviolet spectral range). The probe can be removed and reinsertedinto the container and reused.

FIG. 40 shows an embodiment of a sterilization probe system of theinvention that includes a container which can hold the medium to besterilized, a probe made of material containing an X-ray energyconverter, a dielectric layer (e.g., silica), and a metal manostructure(e.g., Au, Ag). The sample inside the container can be liquid, gas, orparticulates. X-ray radiation, capable of penetrating the containerwall, excites the probe having a material containing X-ray excitationenergy converter (EEC), which in turn emits emission light. The EECmaterial is selected such that the emitted or luminescence light occursin a spectral region that can be used for sterilization (e.g., theultraviolet spectral range). The metal nanostructure is designed toamplify the luminescence light due to the plasmonics enhancement effect.The dielectric layer is designed to separate the material containing theX-ray energy converter and the metal nanostructure in order to preventor minimize possible quenching of the luminescence. The optimalthickness of the dielectric layer is about 1 to 5 nm such that thedielectric layer does not significantly alter the plasmonics effect. Theprobe can be removed and reinserted into the container and reused.

FIG. 41 shows an embodiment of a sterilization probe system of theinvention that includes a container which can hold the medium to besterilized, a probe made of material containing an X-ray energyconverter, and chemical receptors or bioreceptors used to capturetargets. The sample inside the container can be liquid, gas, orparticulates. X-ray radiation, capable of penetrating the containerwall, excites the probe having a material containing X-ray excitationenergy converter (EEC), which in turn emits emission light. The EECmaterial is selected such that the emitted or luminescence light occursin a spectral region that can be used for sterilization (e.g.,ultraviolet spectral range). The layer of chemical receptors (e.g.,ligands specific to chemical groups) or bioreceptors (e.g., antibodies,surface cell receptors) is used to capture biochemical targets ofinterest. In this embodiment, the specific target compounds areselectively bound to the surface of the probe and are more effectivelyirradiated by the emission light. The probe can be removed andreinserted into the container and reused.

FIG. 42 shows an embodiment of a sterilization probe system of theinvention that includes a container which can hold the medium to besterilized, a probe made of material containing an X-ray energyconverter, a dielectric layer (e.g., silica), and a metal manostructure(e.g., Au, Ag). The sample inside the container can be liquid, gas, orparticulates. X-ray radiation, capable of penetrating the containerwall, excites the probe having a material containing X-ray excitationenergy converter (EEC), which in turn emits emission light. The EECmaterial is selected such that the emitted or luminescence light occursin a spectral region that can be used for sterilization (e.g., theultraviolet spectral range). The metal nanostructure is designed toamplify the luminescence light due to the plasmonics enhancement effect.The dielectric layer is designed to separate the material containing theX-ray energy converter and the metal nanostructure in order to preventpossible quenching of the luminescence. The optimal thickness of thedielectric layer s about 1 to 5 nm such that the dielectric layer doesnot significantly affect the plasmonics effect. The layer of chemicalreceptors (e.g., ligands specific to chemical groups) or bioreceptors(e.g., antibodies, surface cell receptors) is used to capturebiochemical targets of interest. In this embodiment, the specific targetcompounds are selectively bound to the surface of the probe and are moreeffectively irradiated by the emission light. The probe can be removedand reinserted into the container and reused.

FIG. 43 shows an embodiment of a sterilization probe system of theinvention that includes a container which can hold the medium to besterilized, nanoparticles having 1) a paramagnetic core and 2) a shellhaving material containing an X-ray energy converter. The sample insidethe container can be liquid, gas, or particulates. The nanoparticles,which have a paramagnetic core covered with a nanoshell of materialcontaining the X-ray energy converter, can be delivered into thecontainer using an externally applied magnetic field. The X-rayradiation, capable of penetrating the container wall, excites thenanoparticle shell which contains X-ray excitation energy converter(EEC) material, which in turn emits emission light. The EEC material isselected such that the emitted or luminescence light occurs in aspectral region that can be used for sterilization (e.g., theultraviolet spectral range). After the sterilization is completed, thenanoparticles can be removed from the container using an externallyapplied magnetic field. The magnetic filed unit serves as a mechanism tointroduce and collect the magnetic naonoparticles. The nanoparticles canbe reinserted into the container and reused. In another embodiment, thenanoparticles can be also covered with a layer of chemical receptors(e.g., ligands specific to chemical groups) or bioreceptors (e.g.,antibodies, surface cell receptors). That layer is used to capturebiochemical targets of interest. In this embodiment, the specific targetcompounds are selectively bound to the surface of the probe and are moreeffectively irradiated by the emission light.

FIG. 44 shows examples of plasmonics probes with a paramagnetic core. inFIG. 44A, the magnetic core is surrounded by a metal layer which is inturn surrounded by a dielectric layer. In FIG. 44B, the magnetic core issurrounded by an X-ray excitation energy converter (EEC) material whichis in turn surrounded by a dielectric layer. Metal nanoparticles areattached to the dielectric. In FIG. 44C, the magnetic core is surroundedby a metal layer which is in turn surrounded by a dielectric layer. AnX-ray excitation energy converter (EEC) material is fornied as a partialcap on the dielectric layer. In FIG. 44D, the magnetic core issurrounded by an X-ray excitation energy converter (EEC) material whichis in turn surrounded by a dielectric layer. A metal layer is formmed asa partial cap on the dielectric layer. In FIG. 44E, the magnetic core issurrounded by a metal layer which is in turn surrounded by a dielectriclayer, which is in turn surrounded by an X-ray excitation energyconverter (EEC) material. In FIG. 44F, the magnetic core is surroundedby an X-ray excitation energy converter (EEC) material which is in turnsurrounded by a dielectric layer, which is in turn surrounded by a metallayer. In FIG. 44G, the magnetic core is surrounded by an X-rayexcitation energy converter (EEC) material which is in turn surroundedby a dielectric layer, which is in turn surrounded by a metal layer andwhich in turn is surrounded by a chemical receptor layer.

Design and Fabrication of Plasmonics-Active Materials and Surfaces

The plasmonics-active surfaces and probes in the embodiments describedabove can be prepared using one of the following procedures to producenanostructures of metal or thin layers of metal that exhibit plasmonicsproperties.

For nanostructures produced on metal electrode systems, electrochemicalcells using silver electrodes and other metal electrodes have been usedto produce nanostructured morphology on the surface of electrodes forSERS studies (Petittinger B., U. Wenneng, and H. Wetzel, Surface-plasmonenhanced Raman-scattering frequency and . . . Ag and Cu electrodes,1980, Surf. Sci., 101, 409; Fleishman M., P. R. Graves, and J. Robinson,The Raman-Spectroscopy of the . . . hydride and platinum-electrodes,1985, J. Electroanal. Chem., 182, 87). The fabrication proceduresdescribed in these references (which are incorporated in their entiretyherein by reference) are applicable to the invention. Silver at anelectrode is oxidized by the reaction Ag→Ag++e− during the first half ofthe cycle. During the reduction half cycle, a roughened silver surfaceis reproduced by the reaction Ag++e−→Ag. This oxidation-reductionprocedure generally produces surface protrusions in the size range of 25to 500 nm on the electrode surface. The working electrode can then begenerally placed in a position such that the laser excitation can befocused onto its surface, and the Raman scattered light can beefficiently collected by appropriate optics. A strong SERS signalsappear in general only after an electrochemical oxidation- reductioncycle, often referred to as “activation cycle,” is performed on themetal electrode. The fabrication procedures described in thesereferences (which are incorporated in their entirety herein byreference) for the respective electrodes are applicable to theinvention.

Other metal electrodes such as platinum (Loo B H., Surface-enhancedRoman-spectroscopy of platinum, 1983, J. Phys. Chem., 87, 3003) havealso been investigated as plasmonics substrates. Experimental factorssuch as the influence of laser illumination of copper electrodes duringoxidation/reduction treatment on SERS signals of pyridine andbenzotriazole have been investigated (Thierry D. and C. Leygraf, Theinfluence of photoalteration nn surface-enhanced. Raman scattering fromcopper electordes, 1985, Surface Sci., 149, 592). Beer, K D.; Tanner,W.; Garrett, R L. in J. Electroanal. Chem. 1989, 258, 313-325. haveinvestigated the ex-situ versus in-situ electrode roughening proceduresfor SERS on gold and silver electrode surfaces. The fabricationprocedures described in these references (which are incorporated intheir entirety herein by reference) for the respective electrodes areapplicable to the invention.

For chemically, electrochemically etched metal and other roughenedsurfaces, chemical etching procedures can also be used to produceplasmonics active metal surfaces (Miller S. K., A. Baiker, M Meier, andA. Wokaun, Surface-enhanced Raman scattering and the preparation ofcopper substrates for catalytic studies, 1984, J. Chem. Soc. Farad.Trans. I, 80, 1305). In one procedure, copper foil is etched for 40 min.in 2 mol. dm⁻³ nitric acid at room temperature. Another procedureincludes sandblasting copper foil with Al₂O₃ at 4 bar pressure andsubsequently etching for 2 min. SEM pictures of the metal surfacesindicate that both etching procedures can produce surface roughness onthe 10 to 100 nm scale. Electrochemically roughened silver oxidesubstrates have been developed to detect vapor of chemical nerve agentsimulants (Taranenko N, J.P. Alarie, D. L. Stokes, and T Vo Dinh,Surface-Enhanced Raman Detection of Nerve Ageiil Sit-111′1(2w (DWP andDIMP) Vapor on Electrochemically Prepared Silver Oxide Substrates, 1996,J. Raman Spectr., 27, 379-384). These procedures are consistent andsimilar to electroplating methods. The fabrication procedures describedin these references (which are incorporated in their entirety herein byreference) are applicable to the invention.

For metallic nanostructures on solid substrates, a variety of proceduresto coat solid substrates with metal nanostructures have been describedpreviously [Vo-Dinh, Suilace-Enhanced Raman Spectroscopy Using MetallicNanostructures, 1998, Trends in Analytical Chemistry, 17,557 (1998)].These procedures can be used to produce plasmonics-active surfaces andembodiments. The fabrication procedures described in this reference(which is incorporated in its entirety herein by reference) areapplicable to the invention.

In various embodiments of the invention, the interior walls can alsohave an appropriate protective coating that is optically transparent tothe emitting light used for sterilization.

For metal nanoparticle island films, the simplest metallic nanostructurecan be produced by evaporating a thin layer (e.g., less than 10 nmthickness) of a metal such as silver directly onto a solid base support.Under these conditions, the silver layer forms nanoparticles on thesupport in the form of isolated metal islands. Upon an increase of thedeposited silver thickness, the particles would start to coalesce andform a continuous film. The size and shape of the metal nanoparticlescan be influenced by varying the thickness of metal deposited (asmeasured by a quartz crystal monitor perpendicular to the evaporationsource). SERS measurements using silver nanoparticle island films werecompared with those obtained with other nanostructures materials. SERSfrom copper and zinc phthalocyanine complexes from silver and indiumisland films were reported (Jennings C., R. Aroca, A. M. Hor, and R. O.Lowly, Surface-enhanced Raman scattering from copper and zincphthalocyanine complexes by silver and indium island films, 1984, Anal.Chem., 56, 203). The silver and indium films were vacuum-evaporated(p<10⁻⁶ Torr) onto tin oxide glass slides and then coated with copperand zinc phthalocyanine complexes in a vacuum system at a base pressureof 5×10⁻⁷ Torr. Metal thickness was about 7.5 nm on the substrates inorder to produce metal nanoparticle islands. Another alternative methodinvolves sputter deposited thin films of metals as plasmonics substrates(Ni F., R. Sheng, and T. M. Cotton, Flow-injection analysis andreal-time . . . bases by surface-enhanced Raman-spectroscopy, 1990,Anal. Chem., 62, 1958). The fabrication procedures described in thesereferences (which are incorporated in their entirety herein byreference) are applicable to the invention.

For metal-coated nanosphere substrates, one of the earlier difficultiesin the development of the SERS technique for analytical applications hadbeen the production of surfaces or media that had an easily controlledprotrusion size (roughness) and reproducible structures. One approachhas involved the use of nanospheres applied onto a solid surface (e.g.,container wall) in order to produce and control the desired roughness.The nanostructured support is subsequently covered with a layer ofsilver that provides the conduction electrons required for the surfaceplasmon mechanisms. Among the techniques based on solid substrates, themethods using simple nanomaterials, such as Teflon or latex nanospheres,appear to be the simplest to prepare. Teflon and latex spheres arecommercially available in a wide variety of sizes. The shapes of thesematerials are very regular and their size can be selected for optimalenhancement. The effect of the sphere size and metal layer thicknesshave indicated that, for each sphere size, there is an optimum silverlayer thickness for which the maximum SERS signal is observed (Moody R.L., T Vo Dinh, and W. H. Fletcher Lniestigation of ExperimentalParameters for Surface-Enhanced Raman Spectroscopy, 1987, Appl. Spectr.,41, 966). The silver-coated nanospheres were found to be among the moststrongly enhancing substrates investigated, with enhancement factorscomparable to or greater than those found for electrochemicallyroughened surfaces. The fabrication procedures described in thisreference (which is incorporated in its entirety herein by reference)are applicable to the invention.

For metal-coated alumina nanoparticles, SERS studies have shown thatnanoparticles with irregular shapes can also be used (instead ofregularly shaped nanospheres) to spin-coat solid substrates. Forinstance, alumina appears to be one of the most efficient materials forthe production of plasmonics-active substrates. The preparation of thesubstrate is similar to that with fumed silica (Bello J. M., D. L.Stokes and T. Vo Dinh, Silver-Coated Aluminum as a New Medium for Slface-Enhanced Raman Scattering Analysis, 1989, Appl. Spectrosc., 43.1325). One important advantage of alumina over Teflon or latexnanospheres is its very low cost. The alumina surface consists ofrandomly distributed surface agglomerates and protrusions in the 10 to100 nm range. These structures produce large electromagnetic fields onthe surface when the incident photon energy is in resonance with thelocalized surface plasmons. Alumina-based substrates, due to theirefficiency, low cost and simplicity for preparation, have led to a widevariety of practical applications. Furthermore, the reproducibility ofalumina-based SERS substrates is excellent; the relative standarddeviation was found to be less than 5% (Sutherland, A PortableSurface-Enhanced Raman Spectrometer, Instrumentation Science &Technology, Volume 22, Issue 3 August 1994 , pages 231 - 239). Thefabrication procedures described in these references (which areincorporated in their entirety herein by reference) are applicable tothe invention.

For silver-coated titanium dioxide nanoparticles, titanium dioxide is analternate material that can be used to produce the nanostnictureroughness when coated on surfaces. The procedures to prepare thesesubstrates are similar to that used for nanospheres and aluminaparticles. Titanium dioxide materials are first deposited on glass andcellulose substrates and then coated with a 50 to 100 nm layer of silverby thermal evaporation as described previously. Prior to deposition,titanium dioxide is prepared as a suspension in water (10% concentrationby weight). The silver-coated titanium oxide surfaces obtained by thismethod provide efficient plasmonics-active substrates (See U.S. Pat. No.7,267,948 whose entire contents are incorporated herein by reference).Titanium dioxide provides the necessary surface nanosized roughness forthe plasmonics effect. Limits of detection of various compounds are inthe part per billion (ppb) levels and demonstrate the analyticalusefulness of this substrate for trace analysis.

For silver-coated silica nanoparticles, another type of substrate thatis quite plasmonics active and easy to prepare is the fumed silica-basedsubstrate (Alak A., and T. Vo Dinh, Silver-Coated Fumed Silica as NewSubstrate Materials lbr Surface-Enhanced Raman Scattering, 1989, Anal.Chem., 61, 656). Fumed silica has been used as a thickening agent invarious industrial processes, including coating and cosmeticspreparations. In the preparation of plasmonics, the selection of theappropriate types of fumed silica is important. Fumed silica ismanufactured in different grades, which vary with respect to surfacearea, particle diameter, and degree of compression. The fumed silicaparticles are suspended in a 10% water solution and coated onto a glassplate or filter paper. The substrate is then coated with a 50 to 100 nmlayer of silver by thermal evaporation. With this type of substrate, thefumed silica material, which has nano-sized structures, provides a roughsurface effect for the plasmonics process. The fabrication proceduresdescribed in this reference (which is incorporated in its entiretyherein by reference) are applicable to the invention.

Plasmonica-active surfaces can be fabricated using lithographictechniques to produce controlled surface roughness have beeninvestigated (Liao P. F., and M. B. Stern, Surface-enhanced Rumorscattering on gold and aluminum particle arrays, 1982, Opt. Lett., 7,483). These surfaces include uniform arrays of isolated silvernanoparticles which are unifolln in shape and size. These surfacesproduce a Raman enhancement on the order of 107 and have been used totest the electromagnetic model of SERS. The effectiveness ofcrossed-grating plasmonics substrates has been compared to that of CaF₂roughened film, island film, and etched quartz (Vo Dinh T., M. Meier,and A. Wokaun, 1986, Surface Enhanced Raman Spectroscopy with SilverParticles on Stochastic Post Substrates, Anal. Chim. Acta, 181, 139).The fabrication procedures described in these references (which areincorporated in their entirety herein by reference) are applicable tothe invention.

Plasma-etched substrates can also be used in the invention. It is oftendifficult to produce periodic structures over large areas bylithographic techniques. The procedure using etched quartz posts avoidsthis difficulty by using an island film as an etch mask on a SiO₂substrate (Enlow P. D., M. C. Buncick, R. J. Warmack, and T. Vo Dinh,Detection of Nitro polynuclear Aromatic Compounds by Surface EnhancedRaman Spectroscopy, 1986, Anal. Chem., 58, 1119). The preparation ofSiO₂ prolate nanorods is a multi-step operation that involves plasmaetching of SiO₂ with a silver island film as an etch mask. Since fusedquartz is etched much more slowly than is thermally deposited quartz, a500 nm layer of SiO₂ is first thermally evaporated onto fused quartz ata rate of 0.1 to 0.2 nm/s. The resulting crystalline quartz is annealedto the fused quartz for 45 min. at approximately 950° C. A 5 nm silverlayer s then evaporated onto the thermal SiO₂ layer and the substrate isflash heated for 20 s at 500° C. This heating causes the thin silverlayer to bead up into small globules, which act as etch masks. Thesubstrate is then etched for 30 to 60 min. in a CHF₃ plasma to producesubmicron prolate SiO₂ posts, which are then coated with a continuous80nm silver layer at normal evaporation angle. Another method includesvarying the angle of evaporation in order to produce silvernanoparticles on the tips of the quartz posts (Vo Dinh T M. Meier, andA. Wokaun, Surface Enhanced Raman Spectroscopy with Silver Particles onStochastic Post Substrates, 1986, Anal. Chim. Acta, 181, 139). Thefabrication procedures described in these references (which areincorporated in their entirety herein by reference) are applicable tothe invention.

Metal-coated cellulose substrates can also be used in the invention.These substrates can be used as (disposable) inner linings ofcontainers. Direct metal coating of special filter papers coated withsilver could provide useful substrates. Certain types of microporefilter papers coated with a thin layer of evaporated silver appear toprovide efficient plasmonics-active substrates. Scanning electronmicrographs of these cellulosic materials have shown that these surfacesconsist of fibrous 10 nm strands with numerous tendrils that provide thenecessary protrusions required for the SERS enhancement.

Silver membranes can also be used in the invention. These membranes canbe also used in the inner lining of containers. One of the simplesttypes of solid substrates is a silver membrane used for air particulatesampling (Vo Dinh T., 1989, Surface-Enhanced Raman Spectrometry, inChemical Analysis of Polycyclic Aromatic Compounds, Wiley, T. Vo-Dinh,Ed., N.Y.). The filter already has nano/micropores and interstices thatprovide the nano/micro features (e.g., nano/micro arrays) required toinduce SERS. Since these membranes, include silver, these membranes canbe used directly as plasmonics-active substrates without necessarilyadding additional silver. The fabrication procedures described in thisreference (which is incorporated in its entirety herein by reference)are applicable to the invention.

There a large variety of micro/nanofabrication techniques that can beused to produce nanostructures on metal substrates. These techniquesinclude (but not limited to) 1) lithography such as for example electronbeam lithography; photolithography, and nanoimprint tithography, 2) dryetching such as for example reactive ion etching (RIE), inductivelycoupled plasma (ICP) etching, and plasma etching, 3) thin filmdeposition and processing, 4) focused ion beam (FEB), 5) e-beam andthermal evaporation, 6) plasma enhanced chemical vapor deposition(PECVD), 7) sputtering, and 8) nanoimprinting

Further, a sol-gel matrix with embedded silver or other metalnanoparticles can also be used in the invention. An opticallytranslucent material has been prepared that acts as a plasmonics-activesubstrate [111. Volcan, D. L. Stokes and T. Vo-Dinh, A Sol-Gel DerivedAgCl Photochromic Coating on Glass for SERS Chemical Sensor Application,Sensors and Actuators B, 106, 660-667 (2005)]. This material is a silicamatrix, synthesized by the sol-gel method and containing in-situprecipitated AgCl particles which serve as precursors for nanoparticlesof elemental silver. Reduction of AgCl to silver nanoparticles isachieved by UV irradiation. The plasmonics-active medium was distributedon solid, hence producing thin, sturdy, and optically translucentsubstrates. This procedure can be further adapted to produce coatingswith embedded metal nanoparticles discussed above. The fabricationprocedures described in this reference (which is incorporated in itsentirety herein by reference) are applicable to the invention.

Having generally described this invention, a further understanding canbe obtained by reference to certain specific examples which are providedherein for purposes of illustration only and are not intended to belimiting unless otherwise specified.

Numerous modifications and variations of the invention are possible inlight of the above teachings. It is therefore to be understood thatwithin the scope of the appended claims, the invention may be practicedotherwise than as specifically described herein.

1.-243. (canceled)
 244. A radiation-cured article, comprising: aradiation-cured medium; and at least one of an energy modulation agentand a plasmonics agent distributed throughout the medium; said energymodulation agent being a substance which converted an initiation energyto a light which cured the radiation-cured medium by polymerization ofpolymers in the radiation-cured medium.
 245. The article of claim 244,wherein said at least one energy modulation agent comprises luminescentagents including at least one of a sulfide, a telluride, a selenide andan oxide semiconductor.
 246. The article of claim 245, wherein theenergy modulation agent comprises at least one of Y₂O₃; ZnS; ZnSe; MgS;CaS; Mn, Er ZnSe; Mn Er MgS; Mn, Er CaS; Mn, Er ZnS; Mn,Yb ZnSe; Mn,YbMgS; Mn, Yb CaS; Mn,Yb ZnS:Tb³⁺, Er³⁺; ZnS:Tb³⁺; Y₂O₃:Tb³⁺; Y₂O₃:Tb³⁺,Er³⁺; ZnS:Mn²⁺; ZnS:Mn,Er³⁺.
 247. The article of claim 245, wherein saidluminescent agents comprise nanotubes, nanoparticles, chemilumiscentparticles, and bioluminescent particles, and mixtures thereof.
 248. Thearticle of claim 245, wherein said luminescent agents comprisesemiconducting or metallic materials.
 249. The article of claim 245,wherein said luminescent agents comprise chemiluminescent agents. 250.The article of claim 244, further comprising color pigments.
 251. Thearticle of claim 244, further comprising fumed silica.
 252. The articleof claim 244, wherein the plasmonics agent comprises metal structures.253. The article of claim 252, wherein the metal structures comprises atleast one of nanospheres, nanorods, nanocubes, nanopyramids, nanoshells,multi-layer nanoshells, and combinations thereof.
 254. The article ofclaim 244, wherein: the energy modulation agent is disposed adjacent toat least one metal nanoparticle serving as the plasmonics agent; theenergy modulation agent is coated at least partially with a metalserving as the plasmonics agent; or the energy modulation agent includesa magnetic substance.
 255. The article of claim 244, wherein: a metalnanoparticle serving as the plasmonics agent is at least partiallycovered with the energy modulation agent; or the metal nanoparticleincludes a magnetic substance.
 256. The article of claim 244, wherein:the plasmonics agent comprises a dielectric-metal nanocomposite; or theplasmonics agent comprises a plurality of differently sized metalnanoparticles disposed in vicinity of each other as a compositeplasmonics agent.
 257. The article of claim 244, wherein the energymodulation agent is configured to receive said initiation energy at ahigher energy than that produced by the energy modulation agent, or at alower energy than that produced by the energy modulation agent.
 258. Aradiation-curable article, comprising: a radiation-curable medium; andat least one energy modulation agent and at least one plasmonics agentdistributed throughout the medium; said energy modulation agent being asubstance which is capable of converting initiation energy to a lightcapable of curing the radiation-curable medium by polymerization ofpolymers in the radiation-curable medium.
 259. The article of claim 258,wherein said energy modulation agent comprises luminescent agentsincluding at least one of a sulfide, a telluride, a selenide and anoxide semiconductor.
 260. The article of claim 261, wherein the energymodulation agent comprises Y₂O₃; ZnS; ZnSe; MgS; CaS; Mn, Er ZnSe; Mn,Er MgS; Mn, Er CaS; Mn, Er ZnS; Mn,Yb ZnSe; Mn,Yb MgS; Mn, Yb CaS; Mn,YbZnS:Tb³⁺, Er³⁺; ZnS:Tb³⁺; Y₂O₃:Tb³⁺; Y₂O₃:Tb³⁺, Er³⁺; ZnS:Mn²⁺;ZnS:Mn,Er³⁺.
 261. The article of claim 259, wherein said luminescentagents comprise nanotubes, nanoparticles, chemilumiscent particles, andbioluminescent particles, and mixtures thereof.
 262. The article ofclaim 259, wherein said luminescent agents comprise nanoparticles ofsemiconducting or metallic materials.
 263. The article of claim 259,wherein said luminescent agents comprise chemiluminescent particles.264. The article of claim 258, further comprising color pigments. 265.The article of claim 258, further comprising fumed silica.
 266. Thearticle of claim 258, wherein the plasmonics agent comprises metalstructures.
 267. The article of claim 266, wherein the metal structurescomprises at least one of nanospheres, nanorods, nanocubes,nanopyramids, nanoshells, multi-layer nanoshells, and combinationsthereof.
 268. The article of claim 258, wherein: the energy modulationagent is disposed adjacent to at least one metal nanoparticle serving asthe plasmonics agent; the energy modulation agent is coated at leastpartially with a metal serving as the plasmonics agent; or the energymodulation agent includes a magnetic substance.
 269. The article ofclaim 258, wherein: a metal nanoparticle serving as the plasmonics agentis at least partially covered with the energy modulation agent; or themetal nanoparticle includes a magnetic substance.
 270. The article ofclaim 258, wherein: the plasmonics agent comprises a dielectric-metalnanocomposite; or the plasmonics agent comprises a plurality ofdifferently sized metal nanoparticles disposed in vicinity of each otheras a composite plasmonics agent.
 271. The article of claim 258, whereinthe energy modulation agent is configured to receive said initiationenergy at a higher energy than that produced by the energy modulationagent or at a lower energy than that produced by the energy modulationagent. 272.-297. (canceled)